Modulators of IRF4 expression

ABSTRACT

The present embodiments provide methods, compounds, and compositions useful for inhibiting IRF4 expression, which may be useful for treating, preventing, or ameliorating a cancer associated with IRF4.

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0332USASEQ_ST25.txt created Aug. 19, 2020, which is 712 kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

FIELD

The present embodiments provide methods, compounds, and compositions useful for inhibiting IRF4 expression, which can be useful for treating, preventing, or ameliorating a cancer associated with IRF4.

BACKGROUND

Interferon Regulatory Factor 4 (IRF4) is a transcription factor involved in immune responses in normal B and T cells, and is strongly implicated in the development of hematological malignancies, especially multiple myeloma (MM). High IRF4 levels is associated with a poor prognosis of overall survival for MM patients. Upregulation of the cereblon/IRF4 pathway accounts for the failure of lenalidomide treatment, an IMiD approved for MM and B cell malignancies. IRF4 is a component of super enhancer in MM cells in which a positive auto-regulatory loop between the oncogene MYC and IRF4 sustains the survival of MM. IRF4 is also involved in cutaneous anaplastic large cell lymphomas DLBCL, B-cell non-Hodgkin's lymphoma, ALL, adult T cell leukemia/lymphoma (ATLL), and peripheral T cell lymphoma. Despite its role in many cancers, IRF4 is considered an undruggable target by conventional therapeutic approaches.

SUMMARY

Certain embodiments provided herein are directed to potent and tolerable compounds and compositions useful for inhibiting IRF4 expression, which can be useful for treating, preventing, ameliorating, or slowing progression of cancer associated with IRF4.

DETAILED DESCRIPTION

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the embodiments, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and/or” unless stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included”, is not limiting.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, treatises, and GenBank and NCBI reference sequence records are hereby expressly incorporated by reference for the portions of the document discussed herein, as well as in their entirety.

It is understood that the sequence set forth in each SEQ ID NO in the examples contained herein is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Compounds described by ION number indicate a combination of nucleobase sequence, chemical modification, and motif.

Unless otherwise indicated, the following terms have the following meanings:

“2′-deoxynucleoside” means a nucleoside comprising 2′-H(H) furanosyl sugar moiety, as found in naturally occurring deoxyribonucleic acids (DNA). In certain embodiments, a 2′-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (uracil).

“2′-O-methoxyethyl” (also 2′-MOE and 2′-O(CH₂)₂—OCH₃) refers to an O-methoxy-ethyl modification at the 2′ position of a furanosyl ring. A 2′-O-methoxyethyl modified sugar is a modified sugar.

“2′-MOE nucleoside” (also 2′-O-methoxyethyl nucleoside) means a nucleoside comprising a 2′-MOE modified sugar moiety.

“2′-substituted nucleoside” or “2-modified nucleoside” means a nucleoside comprising a 2′-substituted or 2′-modified sugar moiety. As used herein, “2′-substituted” or “2-modified” in reference to a sugar moiety means a sugar moiety comprising at least one 2′-substituent group other than H or OH.

“3′ target site” refers to the nucleotide of a target nucleic acid which is complementary to the 3′-most nucleotide of a particular compound.

“5′ target site” refers to the nucleotide of a target nucleic acid which is complementary to the 5′-most nucleotide of a particular compound.

“5-methylcytosine” means a cytosine with a methyl group attached to the 5 position.

“About” means within ±10% of a value. For example, if it is stated, “the compounds affected about 70% inhibition of IRF4”, it is implied that IRF4 levels are inhibited within a range of 60% and 80%.

“Administration” or “administering” refers to routes of introducing a compound or composition provided herein to an individual to perform its intended function. An example of a route of administration that can be used includes, but is not limited to parenteral administration, such as subcutaneous, intravenous, or intramuscular injection or infusion.

“Administered concomitantly” or “co-administration” means administration of two or more compounds in any manner in which the pharmacological effects of both are manifest in the patient. Concomitant administration does not require that both compounds be administered in a single pharmaceutical composition, in the same dosage form, by the same route of administration, or at the same time. The effects of both compounds need not manifest themselves at the same time. The effects need only be overlapping for a period of time and need not be coextensive. Concomitant administration or co-administration encompasses administration in parallel or sequentially.

“Amelioration” refers to an improvement or lessening of at least one indicator, sign, or symptom of an associated disease, disorder, or condition. In certain embodiments, amelioration includes a delay or slowing in the progression or severity of one or more indicators of a condition or disease. The progression or severity of indicators may be determined by subjective or objective measures, which are known to those skilled in the art.

“Animal” refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.

“Antisense activity” means any detectable and/or measurable activity attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the antisense compound to the target.

“Antisense compound” means a compound comprising an oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group. Examples of antisense compounds include single-stranded and double-stranded compounds, such as, oligonucleotides, ribozymes, siRNAs, shRNAs, ssRNAs, and occupancy-based compounds.

“Antisense inhibition” means reduction of target nucleic acid levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels in the absence of the antisense compound.

“Antisense mechanisms” are all those mechanisms involving hybridization of a compound with target nucleic acid, wherein the outcome or effect of the hybridization is either target degradation or target occupancy with concomitant stalling of the cellular machinery involving, for example, transcription or splicing.

“Antisense oligonucleotide” means an oligonucleotide having a nucleobase sequence that is complementary to a target nucleic acid or region or segment thereof. In certain embodiments, an antisense oligonucleotide is specifically hybridizable to a target nucleic acid or region or segment thereof.

“Bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety. “Bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In certain embodiments, the bicyclic sugar moiety does not comprise a furanosyl moiety.

“Branching group” means a group of atoms having at least 3 positions that are capable of forming covalent linkages to at least 3 groups. In certain embodiments, a branching group provides a plurality of reactive sites for connecting tethered ligands to an oligonucleotide via a conjugate linker and/or a cleavable moiety.

“Cell-targeting moiety” means a conjugate group or portion of a conjugate group that is capable of binding to a particular cell type or particular cell types.

“cEt” or “constrained ethyl” means a bicyclic furanosyl sugar moiety comprising a bridge connecting the 4′-carbon and the 2′-carbon, wherein the bridge has the formula: 4′-CH(CH₃)—O-2′.

“cEt nucleoside” means a nucleoside comprising a cEt modified sugar moiety.

“Chemical modification” in a compound describes the substitutions or changes through chemical reaction, of any of the units in the compound relative to the original state of such unit. “Modified nucleoside” means a nucleoside having, independently, a modified sugar moiety and/or modified nucleobase. “Modified oligonucleotide” means an oligonucleotide comprising at least one modified internucleoside linkage, a modified sugar, and/or a modified nucleobase.

“Chemically distinct region” refers to a region of a compound that is in some way chemically different than another region of the same compound. For example, a region having 2′-O-methoxyethyl nucleotides is chemically distinct from a region having nucleotides without 2′-O-methoxyethyl modifications.

“Chimeric antisense compounds” means antisense compounds that have at least 2 chemically distinct regions, each position having a plurality of subunits.

“Chirally enriched population” means a plurality of molecules of identical molecular formula, wherein the number or percentage of molecules within the population that contain a particular stereochemical configuration at a particular chiral center is greater than the number or percentage of molecules expected to contain the same particular stereochemical configuration at the same particular chiral center within the population if the particular chiral center were stereorandom. Chirally enriched populations of molecules having multiple chiral centers within each molecule may contain one or more sterorandom chiral centers. In certain embodiments, the molecules are modified oligonucleotides. In certain embodiments, the molecules are compounds comprising modified oligonucleotides.

“Cleavable bond” means any chemical bond capable of being split. In certain embodiments, a cleavable bond is selected from among: an amide, a polyamide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, a di-sulfide, or a peptide.

“Cleavable moiety” means a bond or group of atoms that is cleaved under physiological conditions, for example, inside a cell, an animal, or a human.

“Complementary” in reference to an oligonucleotide means the nucleobase sequence of such oligonucleotide or one or more regions thereof matches the nucleobase sequence of another oligonucleotide or nucleic acid or one or more regions thereof when the two nucleobase sequences are aligned in opposing directions. Nucleobase matches or complementary nucleobases, as described herein, are limited to the following pairs: adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), and 5-methyl cytosine (^(m)C) and guanine (G) unless otherwise specified. Complementary oligonucleotides and/or nucleic acids need not have nucleobase complementarity at each nucleoside and may include one or more nucleobase mismatches. By contrast, “fully complementary” or “100% complementary” in reference to oligonucleotides means that such oligonucleotides have nucleobase matches at each nucleoside without any nucleobase mismatches.

“Conjugate group” means a group of atoms that is attached to an oligonucleotide. Conjugate groups include a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide.

“Conjugate linker” means a group of atoms comprising at least one bond that connects a conjugate moiety to an oligonucleotide.

“Conjugate moiety” means a group of atoms that is attached to an oligonucleotide via a conjugate linker.

“Contiguous” in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to each other. For example, “contiguous nucleobases” means nucleobases that are immediately adjacent to each other in a sequence.

“Designing” or “Designed to” refer to the process of designing a compound that specifically hybridizes with a selected nucleic acid molecule.

“Diluent” means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, the diluent in an injected composition can be a liquid, e.g. saline solution.

“Differently modified” means chemical modifications or chemical substituents that are different from one another, including absence of modifications. Thus, for example, a MOE nucleoside and an unmodified DNA nucleoside are “differently modified,” even though the DNA nucleoside is unmodified. Likewise, DNA and RNA are “differently modified,” even though both are naturally-occurring unmodified nucleosides. Nucleosides that are the same but for comprising different nucleobases are not differently modified. For example, a nucleoside comprising a 2′-OMe modified sugar and an unmodified adenine nucleobase and a nucleoside comprising a 2′-OMe modified sugar and an unmodified thymine nucleobase are not differently modified.

“Dose” means a specified quantity of a compound or pharmaceutical agent provided in a single administration, or in a specified time period. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose may require a volume not easily accommodated by a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reaction in an individual. In other embodiments, the compound or pharmaceutical agent is administered by infusion over an extended period of time or continuously. Doses may be stated as the amount of pharmaceutical agent per hour, day, week or month.

“Dosing regimen” is a combination of doses designed to achieve one or more desired effects.

“Double-stranded antisense compound” means an antisense compound comprising two oligomeric compounds that are complementary to each other and form a duplex, and wherein one of the two said oligomeric compounds comprises an oligonucleotide.

“Effective amount” means the amount of compound sufficient to effectuate a desired physiological outcome in an individual in need of the compound. The effective amount may vary among individuals depending on the health and physical condition of the individual to be treated, the taxonomic group of the individuals to be treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.

“Efficacy” means the ability to produce a desired effect.

“Expression” includes all the functions by which a gene's coded information is converted into structures present and operating in a cell. Such structures include, but are not limited to, the products of transcription and translation.

“Gapmer” means an oligonucleotide comprising an internal region having a plurality of nucleosides that support RNase H cleavage positioned between external regions having one or more nucleosides, wherein the nucleosides comprising the internal region are chemically distinct from the nucleoside or nucleosides comprising the external regions. The internal region may be referred to as the “gap” and the external regions may be referred to as the “wings.”

“Hybridization” means the annealing of oligonucleotides and/or nucleic acids. While not limited to a particular mechanism, the most common mechanism of hybridization involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a nucleic acid target. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an oligonucleotide and a nucleic acid target.

“Immediately adjacent” means there are no intervening elements between the immediately adjacent elements of the same kind (e.g. no intervening nucleobases between the immediately adjacent nucleobases).

“Individual” means a human or non-human animal selected for treatment or therapy.

“Inhibiting the expression or activity” refers to a reduction or blockade of the expression or activity relative to the expression of activity in an untreated or control sample and does not necessarily indicate a total elimination of expression or activity.

“Internucleoside linkage” means a group or bond that forms a covalent linkage between adjacent nucleosides in an oligonucleotide. “Modified internucleoside linkage” means any internucleoside linkage other than a naturally occurring, phosphate internucleoside linkage. Non-phosphate linkages are referred to herein as modified internucleoside linkages.

“Lengthened oligonucleotides” are those that have one or more additional nucleosides relative to an oligonucleotide disclosed herein, e.g. a parent oligonucleotide.

“Linked nucleosides” means adjacent nucleosides linked together by an internucleoside linkage.

“Linker-nucleoside” means a nucleoside that links an oligonucleotide to a conjugate moiety. Linker-nucleosides are located within the conjugate linker of a compound. Linker-nucleosides are not considered part of the oligonucleotide portion of a compound even if they are contiguous with the oligonucleotide.

“Mismatch” or “non-complementary” means a nucleobase of a first oligonucleotide that is not complementary to the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligonucleotides are aligned. For example, nucleobases including but not limited to a universal nucleobase, inosine, and hypoxanthine, are capable of hybridizing with at least one nucleobase but are still mismatched or non-complementary with respect to nucleobase to which it hybridized. As another example, a nucleobase of a first oligonucleotide that is not capable of hybridizing to the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligonucleotides are aligned is a mismatch or non-complementary nucleobase.

“Modulating” refers to changing or adjusting a feature in a cell, tissue, organ or organism. For example, modulating IRF4 RNA can mean to increase or decrease the level of IRF4 RNA and/or IRF4 protein in a cell, tissue, organ or organism. A “modulator” effects the change in the cell, tissue, organ or organism. For example, a IRF4 compound can be a modulator that decreases the amount of IRF4 RNA and/or IRF4 protein in a cell, tissue, organ or organism.

“MOE” means methoxyethyl.

“Monomer” refers to a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides.

“Motif” means the pattern of unmodified and/or modified sugar moieties, nucleobases, and/or internucleoside linkages, in an oligonucleotide.

“Natural” or “naturally occurring” means found in nature.

“Non-bicyclic modified sugar” or “non-bicyclic modified sugar moiety” means a modified sugar moiety that comprises a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.

“Nucleic acid” refers to molecules composed of monomeric nucleotides. A nucleic acid includes, but is not limited to, ribonucleic acids (RNA), deoxyribonucleic acids (DNA), single-stranded nucleic acids, and double-stranded nucleic acids.

“Nucleobase” means a heterocyclic moiety capable of pairing with a base of another nucleic acid. As used herein a “naturally occurring nucleobase” is adenine (A), thymine (T), cytosine (C), uracil (U), and guanine (G). A “modified nucleobase” is a naturally occurring nucleobase that is chemically modified. A “universal base” or “universal nucleobase” is a nucleobase other than a naturally occurring nucleobase and modified nucleobase, and is capable of pairing with any nucleobase.

“Nucleobase sequence” means the order of contiguous nucleobases in a nucleic acid or oligonucleotide independent of any sugar or internucleoside linkage.

“Nucleoside” means a compound comprising a nucleobase and a sugar moiety. The nucleobase and sugar moiety are each, independently, unmodified or modified. “Modified nucleoside” means a nucleoside comprising a modified nucleobase and/or a modified sugar moiety. Modified nucleosides include abasic nucleosides, which lack a nucleobase.

“Oligomeric compound” means a compound comprising a single oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group.

“Oligonucleotide” means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another. Unless otherwise indicated, oligonucleotides consist of 8-80 linked nucleosides. “Modified oligonucleotide” means an oligonucleotide, wherein at least one sugar, nucleobase, or internucleoside linkage is modified. “Unmodified oligonucleotide” means an oligonucleotide that does not comprise any sugar, nucleobase, or internucleoside modification.

“Parent oligonucleotide” means an oligonucleotide whose sequence is used as the basis of design for more oligonucleotides of similar sequence but with different lengths, motifs, and/or chemistries. The newly designed oligonucleotides may have the same or overlapping sequence as the parent oligonucleotide.

“Parenteral administration” means administration through injection or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g. intrathecal or intracerebroventricular administration.

“Pharmaceutically acceptable carrier or diluent” means any substance suitable for use in administering to an individual. For example, a pharmaceutically acceptable carrier can be a sterile aqueous solution, such as PBS or water-for-injection.

“Pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds, such as oligomeric compounds or oligonucleotides, i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.

“Pharmaceutical agent” means a compound that provides a therapeutic benefit when administered to an individual.

“Pharmaceutical composition” means a mixture of substances suitable for administering to an individual. For example, a pharmaceutical composition may comprise one or more compounds or salt thereof and a sterile aqueous solution.

“Phosphorothioate linkage” means a modified phosphate linkage in which one of the non-bridging oxygen atoms is replaced with a sulfur atom. A phosphorothioate internucleoside linkage is a modified internucleoside linkage.

“Phosphorus moiety” means a group of atoms comprising a phosphorus atom. In certain embodiments, a phosphorus moiety comprises a mono-, di-, or tri-phosphate, or phosphorothioate.

“Portion” means a defined number of contiguous (i.e., linked) nucleobases of a nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an oligomeric compound.

“Prevent” refers to delaying or forestalling the onset, development or progression of a disease, disorder, or condition for a period of time from minutes to indefinitely.

“Prodrug” means a compound in a form outside the body which, when administered to an individual, is metabolized to another form within the body or cells thereof. In certain embodiments, the metabolized form is the active, or more active, form of the compound (e.g., drug). Typically conversion of a prodrug within the body is facilitated by the action of an enzyme(s) (e.g., endogenous or viral enzyme) or chemical(s) present in cells or tissues, and/or by physiologic conditions.

“Reduce” means to bring down to a smaller extent, size, amount, or number.

“RefSeq No.” is a unique combination of letters and numbers assigned to a sequence to indicate the sequence is for a particular target transcript (e.g., target gene). Such sequence and information about the target gene (collectively, the gene record) can be found in a genetic sequence database. Genetic sequence databases include the NCBI Reference Sequence database, GenBank, the European Nucleotide Archive, and the DNA Data Bank of Japan (the latter three forming the International Nucleotide Sequence Database Collaboration or INSDC).

“Region” is defined as a portion of the target nucleic acid having at least one identifiable structure, function, or characteristic.

“RNAi compound” means an antisense compound that acts, at least in part, through RISC or Ago2, but not through RNase H, to modulate a target nucleic acid and/or protein encoded by a target nucleic acid. RNAi compounds include, but are not limited to double-stranded siRNA, single-stranded RNA (ssRNA), and microRNA, including microRNA mimics.

“Segments” are defined as smaller or sub-portions of regions within a nucleic acid.

“Side effects” means physiological disease and/or conditions attributable to a treatment other than the desired effects. In certain embodiments, side effects include injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, myopathies, and malaise. For example, increased aminotransferase levels in serum may indicate liver toxicity or liver function abnormality. For example, increased bilirubin may indicate liver toxicity or liver function abnormality.

“Single-stranded” in reference to a compound means the compound has only one oligonucleotide. “Self-complementary” means an oligonucleotide that at least partially hybridizes to itself. A compound consisting of one oligonucleotide, wherein the oligonucleotide of the compound is self-complementary, is a single-stranded compound. A single-stranded compound may be capable of binding to a complementary compound to form a duplex.

“Sites” are defined as unique nucleobase positions within a target nucleic acid.

“Specifically hybridizable” refers to an oligonucleotide having a sufficient degree of complementarity between the oligonucleotide and a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids. In certain embodiments, specific hybridization occurs under physiological conditions.

“Specifically inhibit” with reference to a target nucleic acid means to reduce or block expression of the target nucleic acid while exhibiting fewer, minimal, or no effects on non-target nucleic acids. Reduction does not necessarily indicate a total elimination of the target nucleic acid's expression.

“Standard cell assay” means assay(s) described in the Examples and reasonable variations thereof.

“Standard in vivo experiment” means the procedure(s) described in the Example(s) and reasonable variations thereof.

“Stereorandom chiral center” in the context of a population of molecules of identical molecular formula means a chiral center having a random stereochemical configuration. For example, in a population of molecules comprising a stereorandom chiral center, the number of molecules having the (S) configuration of the stereorandom chiral center may be but is not necessarily the same as the number of molecules having the (R) configuration of the stereorandom chiral center. The stereochemical configuration of a chiral center is considered random when it is the result of a synthetic method that is not designed to control the stereochemical configuration. In certain embodiments, a stereorandom chiral center is a stereorandom phosphorothioate internucleoside linkage.

“Sugar moiety” means an unmodified sugar moiety or a modified sugar moiety. “Unmodified sugar moiety” or “unmodified sugar” means a 2′-OH(H) furanosyl moiety, as found in RNA (an “unmodified RNA sugar moiety”), or a 2′-H(H) moiety, as found in DNA (an “unmodified DNA sugar moiety”). Unmodified sugar moieties have one hydrogen at each of the 1′, 3′, and 4′ positions, an oxygen at the 3′ position, and two hydrogens at the 5′ position. “Modified sugar moiety” or “modified sugar” means a modified furanosyl sugar moiety or a sugar surrogate. “Modified furanosyl sugar moiety” means a furanosyl sugar comprising a non-hydrogen substituent in place of at least one hydrogen of an unmodified sugar moiety. In certain embodiments, a modified furanosyl sugar moiety is a 2′-substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic sugars and non-bicyclic sugars.

“Sugar surrogate” means a modified sugar moiety having other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, conjugate group, or terminal group in an oligonucleotide. Modified nucleosides comprising sugar surrogates can be incorporated into one or more positions within an oligonucleotide and such oligonucleotides are capable of hybridizing to complementary compounds or nucleic acids.

“Synergy” or “synergize” refers to an effect of a combination that is greater than additive of the effects of each component alone at the same doses.

“IRF4” means any nucleic acid or protein of IRF4. “IRF4 nucleic acid” means any nucleic acid encoding IRF4. For example, in certain embodiments, a IRF4 nucleic acid includes a DNA sequence encoding IRF4, an RNA sequence transcribed from DNA encoding IRF4 (including genomic DNA comprising introns and exons), and an mRNA sequence encoding IRF4. “IRF4 mRNA” means an mRNA encoding a IRF4 protein. The target may be referred to in either upper or lower case.

“IRF4 specific inhibitor” refers to any agent capable of specifically inhibiting IRF4 RNA and/or IRF4 protein expression or activity at the molecular level. For example, IRF4 specific inhibitors include nucleic acids (including antisense compounds), peptides, antibodies, small molecules, and other agents capable of inhibiting the expression of IRF4 RNA and/or IRF4 protein.

“Target gene” refers to a gene encoding a target.

“Targeting” means the specific hybridization of a compound to a target nucleic acid in order to induce a desired effect.

“Target nucleic acid,” “target RNA,” “target RNA transcript” and “nucleic acid target” all mean a nucleic acid capable of being targeted by compounds described herein.

“Target region” means a portion of a target nucleic acid to which one or more compounds is targeted.

“Target segment” means the sequence of nucleotides of a target nucleic acid to which a compound is targeted. “5′ target site” refers to the 5′-most nucleotide of a target segment. “3′ target site” refers to the 3′-most nucleotide of a target segment.

“Terminal group” means a chemical group or group of atoms that is covalently linked to a terminus of an oligonucleotide.

“Therapeutically effective amount” means an amount of a compound, pharmaceutical agent, or composition that provides a therapeutic benefit to an individual.

“Treat” refers to administering a compound or pharmaceutical composition to an animal in order to effect an alteration or improvement of a disease, disorder, or condition in the animal.

CERTAIN EMBODIMENTS

Certain embodiments provide methods, compounds and compositions for inhibiting IRF4 expression.

Certain embodiments provide compounds targeted to a IRF4 nucleic acid. In certain embodiments, the IRF4 nucleic acid has the sequence set forth in RefSeq or GENBANK Accession No. NM_002460.3 or NT_034880.3TRUNC 328000 354000 (incorporated by reference, disclosed herein as SEQ ID NO: 1 and SEQ ID NO: 2, respectively). In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded.

Certain embodiments provide a compound comprising a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide 9 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 9 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide 10 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 10 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide 11 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 11 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 11 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide 12 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 12 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 12 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

Certain embodiments provide a compound comprising a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within the 3′UTR of SEQ ID NO: 1. In certain embodiments, the 3′UTR corresponds to nucleotides 1483 to 5332 of SEQ ID NO: 1. In certain embodiments, a compound comprises a modified oligonucleotide 10 to 30 linked nucleosides in length having a nucleobase sequence at least 85%, at least 90%, at least 95%, or 100% complementary across its entire length to a nucleobase sequence within the 3′UTR of SEQ ID NO: 1. In certain embodiments, the 3′UTR corresponds to nucleotides 1483 to 5332 of SEQ ID NO: 1. In certain embodiments, a compound comprises a modified oligonucleotide 16 to 30 linked nucleosides in length having a nucleobase sequence at least 85%, at least 90%, at least 95%, or 100% complementary across its entire length to a nucleobase sequence within the 3′UTR of SEQ ID NO: 1. In certain embodiments, the 3′UTR corresponds to nucleotides 1483 to 5332 of SEQ ID NO: 1.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 4227-4244, 4227-4242, 4228-4243, or 4229-4244 of SEQ ID NO: 1. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 9667-9682, 11411-11426, or 18090-18105 of SEQ ID NO: 2. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and complementary within nucleotides 4227-4244, 4227-4242, 4228-4243, or 4229-4244 of SEQ ID NO: 1. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and complementary within nucleotides 9667-9682, 11411-11426, or 18090-18105 of SEQ ID NO: 2. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.

In certain embodiments, a compound comprises a modified oligonucleotide 16 linked nucleosides in length having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303.

In certain embodiments, a compound targeted to IRF4 is ION 935918. Out of over 3,000 compounds that were screened as described in the Examples section below, ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834 emerged as the top lead compounds. In particular, ION 935918 exhibited the best combination of properties in terms of potency and tolerability out of over 3,000 compounds.

In certain embodiments, any of the foregoing modified oligonucleotides comprises at least one modified internucleoside linkage, at least one modified sugar, and/or at least one modified nucleobase.

In certain embodiments, any of the foregoing modified oligonucleotides comprises at least one modified sugar. In certain embodiments, at least one modified sugar comprises a 2′-O-methoxyethyl group. In certain embodiments, at least one modified sugar is a bicyclic sugar, such as a 4′-CH(CH₃)—O-2′ group, a 4′-CH₂—O-2′ group, or a 4′-(CH₂)₂—O-2′ group.

In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, such as a phosphorothioate internucleoside linkage.

In certain embodiments, any of the foregoing modified oligonucleotides comprises at least one modified nucleobase, such as 5-methylcytosine.

In certain embodiments, any of the foregoing modified oligonucleotides comprises:

-   -   a gap segment consisting of linked deoxynucleosides;     -   a 5′ wing segment consisting of linked nucleosides; and     -   a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16 to 80 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NO: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length having a nucleobase sequence consisting of the sequence recited in any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 3-3383, wherein the modified oligonucleotide comprises:

-   -   a gap segment consisting of linked deoxynucleosides;     -   a 5′ wing segment consisting of linked nucleosides; and     -   a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303, wherein the modified oligonucleotide comprises:

-   -   a gap segment consisting of linked deoxynucleosides;     -   a 5′ wing segment consisting of linked nucleosides; and     -   a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 1330 or 3303, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of three linked nucleosides; and

a 3′ wing segment consisting of three linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of each wing segment comprises a cEt nucleoside; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 559 or 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of one linked nucleoside; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 1330 or 2021, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of four linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a 2′-O-methoxyethyl nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NO: 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 2021, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 1540, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a 2′-O-methoxyethyl nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In certain embodiments, a compound comprises or consists of ION 935918 or salt thereof, having the following chemical structure:

In certain embodiments, a compound comprises or consists of the sodium salt of ION 935918, having the following chemical structure:

In certain embodiments, a compound comprises or consists of ION 935968 or salt thereof, having the following chemical structure:

In certain embodiments, a compound comprises or consists of the sodium salt of ION 935968, having the following chemical structure:

In any of the foregoing embodiments, the compound or oligonucleotide can be at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to a nucleic acid encoding IRF4.

In any of the foregoing embodiments, the compound can be single-stranded. In certain embodiments, the compound comprises deoxyribonucleotides. In certain embodiments, the compound is double-stranded. In certain embodiments, the compound is double-stranded and comprises ribonucleotides. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

In any of the foregoing embodiments, the compound can be 8 to 80, 10 to 30, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 22, 18 to 24, 18 to 30, 18 to 50, 19 to 22, 19 to 30, 19 to 50, or 20 to 30 linked nucleosides in length. In certain embodiments, the compound comprises or consists of an oligonucleotide.

In certain embodiments, compounds or compositions provided herein comprise a salt of the modified oligonucleotide. In certain embodiments, the salt is a sodium salt. In certain embodiments, the salt is a potassium salt.

In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having at least one of an increase an alanine transaminase (ALT) or aspartate transaminase (AST) value of no more than 4 fold, 3 fold, or 2 fold over saline treated animals or an increase in liver, spleen, or kidney weight of no more than 30%, 20%, 15%, 12%, 10%, 5%, or 2% compared to control treated animals. In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having no increase of ALT or AST over control treated animals. In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having no increase in liver, spleen, or kidney weight over control animals.

Certain embodiments provide a composition comprising the compound of any of the aforementioned embodiments or salt thereof and at least one of a pharmaceutically acceptable carrier or diluent. In certain embodiments, the composition has a viscosity less than about 40 centipoise (cP), less than about 30 centipose (cP), less than about 20 centipose (cP), less than about 15 centipose (cP), or less than about 10 centipose (cP). In certain embodiments, the composition having any of the aforementioned viscosities comprises a compound provided herein at a concentration of about 100 mg/mL, about 125 mg/mL, about 150 mg/mL, about 175 mg/mL, about 200 mg/mL, about 225 mg/mL, about 250 mg/mL, about 275 mg/mL, or about 300 mg/mL. In certain embodiments, the composition having any of the aforementioned viscosities and/or compound concentrations has a temperature of room temperature or about 20° C., about 21° C., about 22° C., about 23° C., about 24° C., about 25° C., about 26° C., about 27° C., about 28° C., about 29° C., or about 30° C.

Certain Indications

Certain embodiments provided herein relate to methods of inhibiting IRF4 expression, which can be useful for treating, preventing, or ameliorating a cancer associated with IRF4 in an individual, by administration of a compound that targets IRF4. In certain embodiments, the compound can be a IRF4 specific inhibitor. In certain embodiments, the compound can be an antisense compound, oligomeric compound, or oligonucleotide targeted to IRF4.

Examples of cancers associated with IRF4 treatable, preventable, and/or ameliorable with the methods provided herein include blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL).

In certain embodiments, a method of treating, preventing, or ameliorating a cancer associated with IRF4 in an individual comprises administering to the individual a compound comprising a IRF4 specific inhibitor, thereby treating, preventing, or ameliorating the cancer. In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, a compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, a compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, a compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, a compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, a compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual parenterally. In certain embodiments, administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.

In certain embodiments, a method of treating or ameliorating caner comprises administering to the individual a compound comprising a IRF4 specific inhibitor, thereby treating or ameliorating the cancer. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual parenterally. In certain embodiments, administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis. In certain embodiments, the individual is identified as having or at risk of having a cancer associated with IRF4.

In certain embodiments, a method of inhibiting expression of IRF4 in an individual having, or at risk of having, a cancer associated with IRF4 comprises administering to the individual a compound comprising a IRF4 specific inhibitor, thereby inhibiting expression of IRF4 in the individual. In certain embodiments, administering the compound inhibits expression of IRF4 in the bone marrow, lymphoid tissue, or lymph node. In certain embodiments, the individual has, or is at risk of having blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual parenterally. In certain embodiments, administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis. In certain embodiments, the individual is identified as having or at risk of having a cancer associated with IRF4.

In certain embodiments, a method of inhibiting expression of IRF4 in a cell comprises contacting the cell with a compound comprising a IRF4 specific inhibitor, thereby inhibiting expression of IRF4 in the cell. In certain embodiments, the cell is a cancer cell. In certain embodiments, the cell is a bone marrow, lymphoid tissue, or lymph node cell. In certain embodiments, the cell is in the bone marrow, lymphoid tissue, or lymph node. In certain embodiments, the cell is in the bone marrow, lymphoid tissue, or lymph node of an individual who has, or is at risk of having cancer, such as blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

In certain embodiments, a method of reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis of an individual having, or at risk of having, a cancer associated with IRF4 comprises administering to the individual a compound comprising a IRF4 specific inhibitor, thereby reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis in the individual. In certain embodiments, the individual has, or is at risk of having, blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. Examples of cancers associated with IRF4 treatable, preventable, and/or ameliorable with the methods provided herein include blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual parenterally. In certain embodiments, the individual is identified as having or at risk of having a cancer associated with IRF4.

Certain embodiments are drawn to a compound comprising a IRF4 specific inhibitor for use in treating cancer. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

Certain embodiments are drawn to a compound comprising a IRF4 specific inhibitor for use in reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis in an individual having cancer. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

Certain embodiments are drawn to use of a compound comprising a IRF4 specific inhibitor for the manufacture or preparation of a medicament for treating cancer. Certain embodiments are drawn to use of a compound comprising a IRF4 specific inhibitor for the preparation of a medicament for treating a cancer associated with IRF4. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma that can be treated with compounds provided herein include, but are not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia that can be treated with compounds provided herein includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

Certain embodiments are drawn to use of a compound comprising a IRF4 specific inhibitor for the manufacture or preparation of a medicament for reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis in an individual having cancer. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. Certain embodiments are drawn to use of a compound comprising a IRF4 specific inhibitor for the preparation of a medicament for reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis in an individual having cancer. In certain embodiments, the cancer is a blood cancer, myeloma, multiple myeloma (MM), B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma includes, but is not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, the compound comprises an antisense compound targeted to IRF4. In certain embodiments, the compound comprises an oligonucleotide targeted to IRF4. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 80 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a modified oligonucleotide of 16 to 80 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.

In any of the foregoing methods or uses, the compound can be targeted to IRF4. In certain embodiments, the compound comprises or consists of a modified oligonucleotide, for example a modified oligonucleotide 8 to 80 linked nucleosides in length, 10 to 30 linked nucleosides in length, 12 to 30 linked nucleosides in length, or 20 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of the nucleobase sequences recited in SEQ ID NOs: 1-2. In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar and/or at least one modified nucleobase. In certain embodiments, the modified internucleoside linkage is a phosphorothioate internucleoside linkage, the modified sugar is a bicyclic sugar or a 2′-O-methoxyethyl, and the modified nucleobase is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide comprises a gap segment consisting of linked deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides, wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In any of the foregoing embodiments, the modified oligonucleotide can be 12 to 30, 15 to 30, 15 to 25, 15 to 24, 16 to 24, 17 to 24, 18 to 24, 19 to 24, 20 to 24, 19 to 22, 20 to 22, 16 to 20, or 17 or 20 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of the nucleobase sequences recited in SEQ ID NOs: 1-2. In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar and/or at least one modified nucleobase. In certain embodiments, the modified internucleoside linkage is a phosphorothioate internucleoside linkage, the modified sugar is a bicyclic sugar or a 2′-O-methoxyethyl, and the modified nucleobase is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide comprises a gap segment consisting of linked 2′-deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides, wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16 to 80 linked nucleosides in length and having a nucleobase sequence comprising any one of SEQ ID NOs: 3-3383, wherein the modified oligonucleotide comprises:

-   -   a gap segment consisting of linked 2′-deoxynucleosides;     -   a 5′ wing segment consisting of linked nucleosides; and     -   a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 2021, 560, 559, 1330, 1540, or 3303, wherein the modified oligonucleotide comprises:

-   -   a gap segment consisting of linked deoxynucleosides;     -   a 5′ wing segment consisting of linked nucleosides; and     -   a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 1330 or 3303, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of three linked nucleosides; and

a 3′ wing segment consisting of three linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of each wing segment comprises a cEt nucleoside; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 559 or 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of one linked nucleoside; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 1330 or 2021, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of four linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a 2′-O-methoxyethyl nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NO: 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 2021, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 1540, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a cEt nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of a modified oligonucleotide 16-80 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in SEQ ID NOs: 560, wherein the modified oligonucleotide comprises:

a gap segment consisting of nine linked deoxynucleosides;

a 5′ wing segment consisting of two linked nucleosides; and

a 3′ wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of the 5′ wing segment comprises a cEt nucleoside; wherein the 3′ wing segment comprises a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, a 2′-O-methoxyethyl nucleoside, a cEt nucleoside, and a 2′-O-methoxyethyl nucleoside in the 5′ to 3′ direction; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.

In any of the foregoing methods or uses, the compound can comprise or consist of ION 935918 or salt thereof, having the following chemical structure:

In any of the foregoing methods or uses, the compound can comprise or consist of the sodium salt of ION 935918, having the following chemical structure:

In any of the foregoing methods or uses, the compound can comprise or consist of ION 935968 or salt thereof, having the following chemical structure:

In any of the foregoing methods or uses, the compound can comprise or consist of the sodium salt of ION 935968, having the following chemical structure:

In any of the foregoing methods or uses, the compound can be administered parenterally. For example, in certain embodiments the compound can be administered through injection or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g. intrathecal or intracerebroventricular administration.

Certain Combinations and Combination Therapies

In certain embodiments, a first agent comprising a compound described herein is co-administered with one or more secondary agents. In certain embodiments, such second agents are designed to treat the same disease, disorder, or condition as the first agent described herein. In certain embodiments, such second agents are designed to treat a different disease, disorder, or condition as the first agent described herein. In certain embodiments, a first agent is designed to treat an undesired side effect of a second agent. In certain embodiments, second agents are co-administered with the first agent to treat an undesired effect of the first agent. In certain embodiments, such second agents are designed to treat an undesired side effect of one or more pharmaceutical compositions as described herein. In certain embodiments, second agents are co-administered with the first agent to produce a combinational effect. In certain embodiments, second agents are co-administered with the first agent to produce a synergistic effect. In certain embodiments, the co-administration of the first and second agents permits use of lower dosages than would be required to achieve a therapeutic or prophylactic effect if the agents were administered as independent therapy.

In certain embodiments, one or more compounds or compositions provided herein are co-administered with one or more secondary agents. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are administered at different times. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are prepared together in a single formulation. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are prepared separately. In certain embodiments, a secondary agent is selected from: proteasome inhibitors including but not limited to bortezomib, carfilzomib, and ixazomib; BTK inhibitors including but not limited to ibrutinib; IMiDs including but not limited to thalidomide, lenalidomide, and pomalidomide; BCL2 inhibitors including but not limited to venetoclax; HDAC inhibitors including but not limited to panobinostat; CDK inhibitors including but not limited to dinaciclib; XPO1 inhibitors including but not limited to selinexor; BET inhibitors including but not limited to CPI-0610; anti-CD38 antibodies including but not limited to daratumumab, isatuximab, and MOR202; anti-CD319 or anti-SLAMF7 antibodies including but not limited to elotuzumab; dexamethasone, cisplatin, doxorubicin, cyclophosphamide, and etoposide.

Certain embodiments are directed to the use of a compound targeted to IRF4 as described herein in combination with a secondary agent. In particular embodiments such use is in a method of treating a patient suffering from cancer including, but not limited to, blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, such use is in the preparation or manufacture of a medicament for treating cancer including, but not limited to, blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia. In certain embodiments, the B-cell lymphoma is a non-Hodgkin's B-cell lymphoma. Examples of non-Hodgkin's B-cell lymphoma of certain embodiments that can be treated with compounds provided herein include, but are not limited to, diffuse large B cell lymphoma (DLBCL), activated B-cell lymphoma (ABC-DLBCL), germinal center B-cell lymphoma (GCB DLBCL), follicular lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), small cell lymphocytic lymphoma, chronic lymphocytic leukemia, mantle cell lymphoma (MCL), Burkitt lymphoma, mediastinal large B cell lymphoma, Waldenström macroglobulinemia, nodal marginal zone B cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. In certain embodiments, the T-cell lymphoma includes, but is not limited to, peripheral T-cell lymphoma, adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL). In certain embodiments, the leukemia includes, but is not limited to, acute lymphocytic leukemia (ALL). In certain embodiments, a secondary agent is selected from: proteasome inhibitors including but not limited to bortezomib, carfilzomib, and ixazomib; BTK inhibitors including but not limited to ibrutinib; IMiDs including but not limited to thalidomide, lenalidomide, and pomalidomide; BCL2 inhibitors including but not limited to venetoclax; HDAC inhibitors including but not limited to panobinostat; CDK inhibitors including but not limited to dinaciclib; XPO1 inhibitors including but not limited to selinexor; BET inhibitors including but not limited to CPI-0610; anti-CD38 antibodies including but not limited to daratumumab, isatuximab, and MOR202; anti-CD319 or anti-SLAMF7 antibodies including but not limited to elotuzumab; dexamethasone, cisplatin, doxorubicin, cyclophosphamide, and etoposide.

Certain embodiments are drawn to a combination of a compound targeted to IRF4 as described herein and a secondary agent, such as a secondary agent selected from: proteasome inhibitors including but not limited to bortezomib, carfilzomib, and ixazomib; BTK inhibitors including but not limited to ibrutinib; IMiDs including but not limited to thalidomide, lenalidomide, and pomalidomide; BCL2 inhibitors including but not limited to venetoclax; HDAC inhibitors including but not limited to panobinostat; CDK inhibitors including but not limited to dinaciclib; XPO1 inhibitors including but not limited to selinexor; BET inhibitors including but not limited to CPI-0610; dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide. In certain embodiments, such a combination of a compound targeted to IRF4 as described herein and a secondary agent, such as a secondary agent selected from: proteasome inhibitors including but not limited to bortezomib, carfilzomib, and ixazomib; BTK inhibitors including but not limited to ibrutinib; IMiDs including but not limited to lenalidomide; BCL2 inhibitors including but not limited to venetoclax; HDAC inhibitors including but not limited to panobinostat; CDK inhibitors including but not limited to dinaciclib; XPO1 inhibitors including but not limited to selinexor; BET inhibitors including but not limited to CPI-0610; anti-CD38 antibodies including but not limited to daratumumab, isatuximab, and MOR202; anti-CD319 or anti-SLAMF7 antibodies including but not limited to elotuzumab; dexamethasone, cisplatin, doxorubicin, cyclophosphamide, and etoposide. Such combinations can be useful for reducing or inhibiting cancer cell proliferation, tumor growth, or metastasis and/or treating cancer including, but not limited to, blood cancer, myeloma, multiple myeloma (MM), B cell malignancies, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia.

In certain embodiments the compound targeted to IRF4 as described herein and the secondary agent are used in combination treatment by administering the two agents simultaneously, separately or sequentially. In certain embodiments the two agents are formulated as a fixed dose combination product. In other embodiments the two agents are provided to the patient as separate units which can then either be taken simultaneously or serially (sequentially).

Certain Compounds

In certain embodiments, compounds described herein can be antisense compounds. In certain embodiments, the antisense compound comprises or consists of an oligomeric compound. In certain embodiments, the oligomeric compound comprises a modified oligonucleotide. In certain embodiments, the modified oligonucleotide has a nucleobase sequence complementary to that of a target nucleic acid.

In certain embodiments, a compound described herein comprises or consists of a modified oligonucleotide. In certain embodiments, the modified oligonucleotide has a nucleobase sequence complementary to that of a target nucleic acid.

In certain embodiments, a compound or antisense compound is single-stranded. Such a single-stranded compound or antisense compound comprises or consists of an oligomeric compound. In certain embodiments, such an oligomeric compound comprises or consists of an oligonucleotide and optionally a conjugate group. In certain embodiments, the oligonucleotide is an antisense oligonucleotide. In certain embodiments, the oligonucleotide is modified. In certain embodiments, the oligonucleotide of a single-stranded antisense compound or oligomeric compound comprises a self-complementary nucleobase sequence.

In certain embodiments, compounds are double-stranded. Such double-stranded compounds comprise a first modified oligonucleotide having a region complementary to a target nucleic acid and a second modified oligonucleotide having a region complementary to the first modified oligonucleotide. In certain embodiments, the modified oligonucleotide is an RNA oligonucleotide. In such embodiments, the thymine nucleobase in the modified oligonucleotide is replaced by a uracil nucleobase. In certain embodiments, compound comprises a conjugate group. In certain embodiments, one of the modified oligonucleotides is conjugated. In certain embodiments, both the modified oligonucleotides are conjugated. In certain embodiments, the first modified oligonucleotide is conjugated. In certain embodiments, the second modified oligonucleotide is conjugated. In certain embodiments, the first modified oligonucleotide is 12-30 linked nucleosides in length and the second modified oligonucleotide is 12-30 linked nucleosides in length. In certain embodiments, one of the modified oligonucleotides has a nucleobase sequence comprising at least 8 contiguous nucleobases of any of SEQ ID NOs: 3-3383.

In certain embodiments, antisense compounds are double-stranded. Such double-stranded antisense compounds comprise a first oligomeric compound having a region complementary to a target nucleic acid and a second oligomeric compound having a region complementary to the first oligomeric compound. The first oligomeric compound of such double stranded antisense compounds typically comprises or consists of a modified oligonucleotide and optionally a conjugate group. The oligonucleotide of the second oligomeric compound of such double-stranded antisense compound may be modified or unmodified. Either or both oligomeric compounds of a double-stranded antisense compound may comprise a conjugate group. The oligomeric compounds of double-stranded antisense compounds may include non-complementary overhanging nucleosides.

Examples of single-stranded and double-stranded compounds include but are not limited to oligonucleotides, siRNAs, microRNA targeting oligonucleotides, and single-stranded RNAi compounds, such as small hairpin RNAs (shRNAs), single-stranded siRNAs (ssRNAs), and microRNA mimics.

In certain embodiments, a compound described herein has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.

In certain embodiments, a compound described herein comprises an oligonucleotide 10 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 12 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 12 to 22 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 14 to 30 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 14 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 15 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 15 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 21 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 20 to 30 linked subunits in length. In other words, such oligonucleotides are 12 to 30 linked subunits, 14 to 30 linked subunits, 14 to 20 subunits, 15 to 30 subunits, 15 to 20 subunits, 16 to 30 subunits, 16 to 20 subunits, 17 to 30 subunits, 17 to 20 subunits, 18 to 30 subunits, 18 to 20 subunits, 18 to 21 subunits, 20 to 30 subunits, or 12 to 22 linked subunits in length, respectively. In certain embodiments, a compound described herein comprises an oligonucleotide 14 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 18 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 19 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 20 linked subunits in length. In other embodiments, a compound described herein comprises an oligonucleotide 8 to 80, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 22, 18 to 24, 18 to 30, 18 to 50, 19 to 22, 19 to 30, 19 to 50, or 20 to 30 linked subunits. In certain such embodiments, the compound described herein comprises an oligonucleotide 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 linked subunits in length, or a range defined by any two of the above values. In some embodiments the linked subunits are nucleotides, nucleosides, or nucleobases.

In certain embodiments, the compound may further comprise additional features or elements, such as a conjugate group, that are attached to the oligonucleotide. In certain embodiments, such compounds are antisense compounds. In certain embodiments, such compounds are oligomeric compounds. In embodiments where a conjugate group comprises a nucleoside (i.e. a nucleoside that links the conjugate group to the oligonucleotide), the nucleoside of the conjugate group is not counted in the length of the oligonucleotide.

In certain embodiments, compounds may be shortened or truncated. For example, a single subunit may be deleted from the 5′ end (5′ truncation), or alternatively from the 3′ end (3′ truncation). A shortened or truncated compound targeted to an IRF4 nucleic acid may have two subunits deleted from the 5′ end, or alternatively may have two subunits deleted from the 3′ end, of the compound. Alternatively, the deleted nucleosides may be dispersed throughout the compound.

When a single additional subunit is present in a lengthened compound, the additional subunit may be located at the 5′ or 3′ end of the compound. When two or more additional subunits are present, the added subunits may be adjacent to each other, for example, in a compound having two subunits added to the 5′ end (5′ addition), or alternatively to the 3′ end (3′ addition), of the compound. Alternatively, the added subunits may be dispersed throughout the compound.

It is possible to increase or decrease the length of a compound, such as an oligonucleotide, and/or introduce mismatch bases without eliminating activity (Woolf et al. Proc. Natl. Acad. Sci. USA 1992, 89:7305-7309; Gautschi et al. J. Natl. Cancer Inst. March 2001, 93:463-471; Maher and Dolnick Nuc. Acid. Res. 1998, 16:3341-3358). However, seemingly small changes in oligonucleotide sequence, chemistry and motif can make large differences in one or more of the many properties required for clinical development (Seth et al. J. Med. Chem. 2009, 52, 10; Egli et al. J. Am. Chem. Soc. 2011, 133, 16642).

In certain embodiments, compounds described herein are interfering RNA compounds (RNAi), which include double-stranded RNA compounds (also referred to as short-interfering RNA or siRNA) and single-stranded RNAi compounds (or ssRNA). Such compounds work at least in part through the RISC pathway to degrade and/or sequester a target nucleic acid (thus, include microRNA/microRNA-mimic compounds). As used herein, the term siRNA is meant to be equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid, short interfering modified oligonucleotide, chemically modified siRNA, post-transcriptional gene silencing RNA (ptgsRNA), and others. In addition, as used herein, the term “RNAi” is meant to be equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, or epigenetics.

In certain embodiments, a compound described herein can comprise any of the oligonucleotide sequences targeted to IRF4 described herein. In certain embodiments, the compound can be double-stranded. In certain embodiments, the compound comprises a first strand comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 3-3383 and a second strand. In certain embodiments, the compound comprises a first strand comprising the nucleobase sequence of any one of SEQ ID NOs: 3-3383 and a second strand. In certain embodiments, the compound comprises ribonucleotides in which the first strand has uracil (U) in place of thymine (T) in any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises (i) a first strand comprising a nucleobase sequence complementary to the site on IRF4 to which any of SEQ ID NOs: 3-3383 is targeted, and (ii) a second strand. In certain embodiments, the compound comprises one or more modified nucleotides in which the 2′ position in the sugar contains a halogen (such as fluorine group; 2′-F) or contains an alkoxy group (such as a methoxy group; 2′-OMe). In certain embodiments, the compound comprises at least one 2′-F sugar modification and at least one 2′-OMe sugar modification. In certain embodiments, the at least one 2′-F sugar modification and at least one 2′-OMe sugar modification are arranged in an alternating pattern for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases along a strand of the dsRNA compound. In certain embodiments, the compound comprises one or more linkages between adjacent nucleotides other than a naturally-occurring phosphodiester linkage. Examples of such linkages include phosphoramide, phosphorothioate, and phosphorodithioate linkages. The compounds may also be chemically modified nucleic acid molecules as taught in U.S. Pat. No. 6,673,661. In other embodiments, the compound contains one or two capped strands, as disclosed, for example, by WO 00/63364, filed Apr. 19, 2000.

In certain embodiments, the first strand of the compound is an siRNA guide strand and the second strand of the compound is an siRNA passenger strand. In certain embodiments, the second strand of the compound is complementary to the first strand. In certain embodiments, each strand of the compound is 16, 17, 18, 19, 20, 21, 22, or 23 linked nucleosides in length. In certain embodiments, the first or second strand of the compound can comprise a conjugate group.

In certain embodiments, a compound described herein can comprise any of the oligonucleotide sequences targeted to IRF4 described herein. In certain embodiments, the compound is single stranded. In certain embodiments, such a compound is a single-stranded RNAi (ssRNAi) compound. In certain embodiments, the compound comprises at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises the nucleobase sequence of any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises ribonucleotides in which uracil (U) is in place of thymine (T) in any one of SEQ ID NOs: 3-3383. In certain embodiments, the compound comprises a nucleobase sequence complementary to the site on IRF4 to which any of SEQ ID NOs: 3-3383 is targeted. In certain embodiments, the compound comprises one or more modified nucleotides in which the 2′ position in the sugar contains a halogen (such as fluorine group; 2′-F) or contains an alkoxy group (such as a methoxy group; 2′-OMe). In certain embodiments, the compound comprises at least one 2′-F sugar modification and at least one 2′-OMe sugar modification. In certain embodiments, the at least one 2′-F sugar modification and at least one 2′-OMe sugar modification are arranged in an alternating pattern for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases along a strand of the compound. In certain embodiments, the compound comprises one or more linkages between adjacent nucleotides other than a naturally-occurring phosphodiester linkage. Examples of such linkages include phosphoramide, phosphorothioate, and phosphorodithioate linkages. The compounds may also be chemically modified nucleic acid molecules as taught in U.S. Pat. No. 6,673,661. In other embodiments, the compound contains a capped strand, as disclosed, for example, by WO 00/63364, filed Apr. 19, 2000. In certain embodiments, the compound consists of 16, 17, 18, 19, 20, 21, 22, or 23 linked nucleosides. In certain embodiments, the compound can comprise a conjugate group.

In certain embodiments, compounds described herein comprise modified oligonucleotides. Certain modified oligonucleotides have one or more asymmetric center and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), as α or β such as for sugar anomers, or as (D) or (L) such as for amino acids etc. Included in the modified oligonucleotides provided herein are all such possible isomers, including their racemic and optically pure forms, unless specified otherwise. Likewise, all cis- and trans-isomers and tautomeric forms are also included.

The compounds described herein include variations in which one or more atoms are replaced with a non-radioactive isotope or radioactive isotope of the indicated element. For example, compounds herein that comprise hydrogen atoms encompass all possible deuterium substitutions for each of the ¹H hydrogen atoms. Isotopic substitutions encompassed by the compounds herein include but are not limited to: ²H or ³H in place of ¹H, ¹³C or ¹⁴C in place of ¹²C, ¹⁵N in place of ¹⁴N, ¹⁷O or ¹⁸O in place of ¹⁶O, and ³³S, ³⁴S, ³⁵S, or ³⁶S in place of ³²S. In certain embodiments, non-radioactive isotopic substitutions may impart new properties on the compound that are beneficial for use as a therapeutic or research tool. In certain embodiments, radioactive isotopic substitutions may make the compound suitable for research or diagnostic purposes, such as an imaging assay.

Certain Mechanisms

In certain embodiments, compounds described herein comprise or consist of modified oligonucleotides. In certain embodiments, compounds described herein are antisense compounds. In certain embodiments, compounds comprise oligomeric compounds. In certain embodiments, compounds described herein are capable of hybridizing to a target nucleic acid, resulting in at least one antisense activity. In certain embodiments, compounds described herein selectively affect one or more target nucleic acid. Such compounds comprise a nucleobase sequence that hybridizes to one or more target nucleic acid, resulting in one or more desired antisense activity and does not hybridize to one or more non-target nucleic acid or does not hybridize to one or more non-target nucleic acid in such a way that results in a significant undesired antisense activity.

In certain antisense activities, hybridization of a compound described herein to a target nucleic acid results in recruitment of a protein that cleaves the target nucleic acid. For example, certain compounds described herein result in RNase H mediated cleavage of the target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not be unmodified DNA. In certain embodiments, compounds described herein are sufficiently “DNA-like” to elicit RNase H activity. Further, in certain embodiments, one or more non-DNA-like nucleoside in the gap of a gapmer is tolerated.

In certain antisense activities, compounds described herein or a portion of the compound is loaded into an RNA-induced silencing complex (RISC), ultimately resulting in cleavage of the target nucleic acid. For example, certain compounds described herein result in cleavage of the target nucleic acid by Argonaute. Compounds that are loaded into RISC are RNAi compounds. RNAi compounds may be double-stranded (siRNA) or single-stranded (ssRNA).

In certain embodiments, hybridization of compounds described herein to a target nucleic acid does not result in recruitment of a protein that cleaves that target nucleic acid. In certain such embodiments, hybridization of the compound to the target nucleic acid results in alteration of splicing of the target nucleic acid. In certain embodiments, hybridization of the compound to a target nucleic acid results in inhibition of a binding interaction between the target nucleic acid and a protein or other nucleic acid. In certain such embodiments, hybridization of the compound to a target nucleic acid results in alteration of translation of the target nucleic acid.

Antisense activities may be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid, a change in the ratio of splice variants of a nucleic acid or protein, and/or a phenotypic change in a cell or animal.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

In certain embodiments, compounds described herein comprise or consist of an oligonucleotide comprising a region that is complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the target nucleic acid encodes a protein. In certain such embodiments, the target nucleic acid is selected from: an mRNA and a pre-mRNA, including intronic, exonic and untranslated regions. In certain embodiments, the target RNA is an mRNA. In certain embodiments, the target nucleic acid is a pre-mRNA. In certain such embodiments, the target region is entirely within an intron. In certain embodiments, the target region spans an intron/exon junction. In certain embodiments, the target region is at least 50% within an intron.

Nucleotide sequences that encode IRF4 include, without limitation, the following: RefSEQ No. NM_002460.3 and NT_034880.3_TRUNC_328000_354000.

Hybridization

In some embodiments, hybridization occurs between a compound disclosed herein and a IRF4 nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.

Hybridization can occur under varying conditions. Hybridization conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

Methods of determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In certain embodiments, the compounds provided herein are specifically hybridizable with a IRF4 nucleic acid.

Complementarity

An oligonucleotide is said to be complementary to another nucleic acid when the nucleobase sequence of such oligonucleotide or one or more regions thereof matches the nucleobase sequence of another oligonucleotide or nucleic acid or one or more regions thereof when the two nucleobase sequences are aligned in opposing directions. Nucleobase matches or complementary nucleobases, as described herein, are limited to the following pairs: adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), and 5-methyl cytosine (mC) and guanine (G) unless otherwise specified. Complementary oligonucleotides and/or nucleic acids need not have nucleobase complementarity at each nucleoside and may include one or more nucleobase mismatches. An oligonucleotide is fully complementary or 100% complementary when such oligonucleotides have nucleobase matches at each nucleoside without any nucleobase mismatches.

In certain embodiments, compounds described herein comprise or consist of modified oligonucleotides. In certain embodiments, compounds described herein are antisense compounds. In certain embodiments, compounds comprise oligomeric compounds. Non-complementary nucleobases between a compound and a IRF4 nucleic acid may be tolerated provided that the compound remains able to specifically hybridize to a target nucleic acid. Moreover, a compound may hybridize over one or more segments of a IRF4 nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).

In certain embodiments, the compounds provided herein, or a specified portion thereof, are, are at least, or are up to 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a IRF4 nucleic acid, a target region, target segment, or specified portion thereof. In certain embodiments, the compounds provided herein, or a specified portion thereof, are 70% to 75%, 75% to 80%, 80% to 85%, 85% to 90%, 90% to 95%, 95% to 100%, or any number in between these ranges, complementary to a IRF4 nucleic acid, a target region, target segment, or specified portion thereof. Percent complementarity of a compound with a target nucleic acid can be determined using routine methods.

For example, a compound in which 18 of 20 nucleobases of the compound are complementary to a target region, and would therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining non-complementary nucleobases may be clustered or interspersed with complementary nucleobases and need not be contiguous to each other or to complementary nucleobases. As such, a compound which is 18 nucleobases in length having four non-complementary nucleobases which are flanked by two regions of complete complementarity with the target nucleic acid would have 77.8% overall complementarity with the target nucleic acid. Percent complementarity of a compound with a region of a target nucleic acid can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403 410; Zhang and Madden, Genome Res., 1997, 7, 649 656). Percent homology, sequence identity or complementarity, can be determined by, for example, the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which uses the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482 489).

In certain embodiments, compounds described herein, or specified portions thereof, are fully complementary (i.e. 100% complementary) to a target nucleic acid, or specified portion thereof. For example, a compound may be fully complementary to a IRF4 nucleic acid, or a target region, or a target segment or target sequence thereof. As used herein, “fully complementary” means each nucleobase of a compound is complementary to the corresponding nucleobase of a target nucleic acid. For example, a 20 nucleobase compound is fully complementary to a target sequence that is 400 nucleobases long, so long as there is a corresponding 20 nucleobase portion of the target nucleic acid that is fully complementary to the compound. Fully complementary can also be used in reference to a specified portion of the first and/or the second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase compound can be “fully complementary” to a target sequence that is 400 nucleobases long. The 20 nucleobase portion of the 30 nucleobase compound is fully complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion wherein each nucleobase is complementary to the 20 nucleobase portion of the compound. At the same time, the entire 30 nucleobase compound may or may not be fully complementary to the target sequence, depending on whether the remaining 10 nucleobases of the compound are also complementary to the target sequence.

In certain embodiments, compounds described herein comprise one or more mismatched nucleobases relative to the target nucleic acid. In certain such embodiments, antisense activity against the target is reduced by such mismatch, but activity against a non-target is reduced by a greater amount. Thus, in certain such embodiments selectivity of the compound is improved. In certain embodiments, the mismatch is specifically positioned within an oligonucleotide having a gapmer motif. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5′-end of the gap region. In certain such embodiments, the mismatch is at position 9, 8, 7, 6, 5, 4, 3, 2, 1 from the 3′-end of the gap region. In certain such embodiments, the mismatch is at position 1, 2, 3, or 4 from the 5′-end of the wing region. In certain such embodiments, the mismatch is at position 4, 3, 2, or 1 from the 3′-end of the wing region. In certain embodiments, the mismatch is specifically positioned within an oligonucleotide not having a gapmer motif. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 5′-end of the oligonucleotide. In certain such embodiments, the mismatch is at position, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 3′-end of the oligonucleotide.

The location of a non-complementary nucleobase may be at the 5′ end or 3′ end of the compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e. linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer oligonucleotide.

In certain embodiments, compounds described herein that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleobases in length comprise no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a IRF4 nucleic acid, or specified portion thereof.

In certain embodiments, compounds described herein that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a IRF4 nucleic acid, or specified portion thereof.

In certain embodiments, compounds described herein also include those which are complementary to a portion of a target nucleic acid. As used herein, “portion” refers to a defined number of contiguous (i.e. linked) nucleobases within a region or segment of a target nucleic acid. A “portion” can also refer to a defined number of contiguous nucleobases of a compound. In certain embodiments, the-compounds, are complementary to at least an 8 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 9 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 10 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least an 11 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 12 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 13 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 14 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 15 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 16 nucleobase portion of a target segment. Also contemplated are compounds that are complementary to at least a 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more nucleobase portion of a target segment, or a range defined by any two of these values.

Identity

The compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific ION number, or portion thereof. In certain embodiments, compounds described herein are antisense compounds or oligomeric compounds. In certain embodiments, compounds described herein are modified oligonucleotides. As used herein, a compound is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in a disclosed DNA sequence would be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the compounds described herein as well as compounds having non-identical bases relative to the compounds provided herein also are contemplated. The non-identical bases may be adjacent to each other or dispersed throughout the compound. Percent identity of an compound is calculated according to the number of bases that have identical base pairing relative to the sequence to which it is being compared.

In certain embodiments, compounds described herein, or portions thereof, are, or are at least, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to one or more of the compounds or SEQ ID NOs, or a portion thereof, disclosed herein. In certain embodiments, compounds described herein are about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, or any percentage between such values, to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific ION number, or portion thereof, in which the compounds comprise an oligonucleotide having one or more mismatched nucleobases. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 5′-end of the oligonucleotide. In certain such embodiments, the mismatch is at position, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 3′-end of the oligonucleotide.

In certain embodiments, compounds described herein comprise or consist of antisense compounds. In certain embodiments, a portion of the antisense compound is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

In certain embodiments, compounds described herein comprise or consist of oligonucleotides. In certain embodiments, a portion of the oligonucleotide is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

Certain Modified Compounds

In certain embodiments, compounds described herein comprise or consist of oligonucleotides consisting of linked nucleosides. Oligonucleotides may be unmodified oligonucleotides (RNA or DNA) or may be modified oligonucleotides. Modified oligonucleotides comprise at least one modification relative to unmodified RNA or DNA (i.e., comprise at least one modified nucleoside (comprising a modified sugar moiety and/or a modified nucleobase) and/or at least one modified internucleoside linkage).

A. Modified Nucleosides

Modified nucleosides comprise a modified sugar moiety or a modified nucleobase or both a modified sugar moiety and a modified nucleobase.

1. Modified Sugar Moieties

In certain embodiments, sugar moieties are non-bicyclic modified sugar moieties. In certain embodiments, modified sugar moieties are bicyclic or tricyclic sugar moieties. In certain embodiments, modified sugar moieties are sugar surrogates. Such sugar surrogates may comprise one or more substitutions corresponding to those of other types of modified sugar moieties.

In certain embodiments, modified sugar moieties are non-bicyclic modified sugar moieties comprising a furanosyl ring with one or more acyclic substituent, including but not limited to substituents at the 2′, 4′, and/or 5′ positions. In certain embodiments one or more acyclic substituent of non-bicyclic modified sugar moieties is branched. Examples of 2′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 2′-F, 2′-OCH₃-(“OMe” or “O-methyl”), and 2′-O(CH₂)₂OCH₃ (“MOE”). In certain embodiments, 2′-substituent groups are selected from among: halo, allyl, amino, azido, SH, CN, OCN, CF₃, OCF₃, alkoxy, O—C₁-C₁₀ substituted alkoxy, O—C₁-C₁₀ alkyl, O—C₁-C₁₀ substituted alkyl, S-alkyl, N(R_(m))-alkyl, O-alkenyl, S-alkenyl, N(R_(m))-alkenyl, O-alkynyl, S-alkynyl, N(R_(m))-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH₂)₂SCH₃, O(CH₂)₂ON(R_(m))(R_(n)) or OCH₂C(═O)—N(R_(m))(R_(n)), where each R_(m) and R_(n) is, independently, H, an amino protecting group, or substituted or unsubstituted C₁-C₁₀ alkyl, and the 2′-substituent groups described in Cook et al., U.S. Pat. No. 6,531,584; Cook et al., U.S. Pat. No. 5,859,221; and Cook et al., U.S. Pat. No. 6,005,087. Certain embodiments of these 2′-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO₂), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. Examples of 4′-substituent groups suitable for linearly non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015/106128. Examples of 5′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 5′-methyl (R or S), 5′-vinyl, and 5′-methoxy. In certain embodiments, non-bicyclic modified sugars comprise more than one non-bridging sugar substituent, for example, 2′-F-5′-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., US2010/190837 and Rajeev et al., US2013/0203836.

In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a linear 2′-substituent group selected from: F, NH₂, N₃, OCF₃, OCH₃, O(CH₂)₃NH₂, CH₂CH═CH₂, OCH₂CH═CH₂, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃, O(CH₂)₂ON(R_(m))(R_(n)), O(CH₂)₂O(CH₂)₂N(CH₃)₂, and N-substituted acetamide (OCH₂C(═O)—N(R_(m))(R_(n))), where each R_(m) and R_(n) is, independently, H, an amino protecting group, or substituted or unsubstituted C₁-C₁₀ alkyl.

In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a linear 2′-substituent group selected from: F, OCF₃, OCH₃, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃, O(CH₂)₂ON(CH₃)₂, O(CH₂)₂O(CH₂)₂N(CH₃)₂, and OCH₂C(═O)—N(H)CH₃ (“NMA”).

In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a linear 2′-substituent group selected from: F, OCH₃, and OCH₂CH₂OCH₃.

Nucleosides comprising modified sugar moieties, such as non-bicyclic modified sugar moieties, are referred to by the position(s) of the substitution(s) on the sugar moiety of the nucleoside. For example, nucleosides comprising 2′-substituted or 2-modified sugar moieties are referred to as 2′-substituted nucleosides or 2-modified nucleosides.

Certain modified sugar moieties comprise a bridging sugar substituent that forms a second ring resulting in a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety comprises a bridge between the 4′ and the 2′ furanose ring atoms. Examples of such 4′ to 2′ bridging sugar substituents include but are not limited to: 4′-CH₂-2′, 4′-(CH₂)₂-2′, 4′-(CH₂)₃-2′, 4′-CH₂—O-2′ (“LNA”), 4′-CH₂—S-2′, 4′-(CH₂)₂—O-2′ (“ENA”), 4′-CH(CH₃)—O-2′ (referred to as “constrained ethyl” or “cEt” when in the S configuration), 4′-CH₂—O—CH₂-2′, 4′-CH₂—N(R)-2′, 4′-CH(CH₂OCH₃)—O-2′ (“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 7,399,845, Bhat et al., U.S. Pat. No. 7,569,686, Swayze et al., U.S. Pat. No. 7,741,457, and Swayze et al., U.S. Pat. No. 8,022,193), 4′-C(CH₃)(CH₃)—O-2′ and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 8,278,283), 4′-CH₂—N(OCH₃)-2′ and analogs thereof (see, e.g., Prakash et al., U.S. Pat. No. 8,278,425), 4′-CH₂—O—N(CH₃)-2′ (see, e.g., Allerson et al., U.S. Pat. No. 7,696,345 and Allerson et al., U.S. Pat. No. 8,124,745), 4′-CH₂—C(H)(CH₃)-2′ (see, e.g., Zhou, et al., J. Org. Chem., 2009, 74, 118-134), 4′-CH₂—C(═CH₂)-2′ and analogs thereof (see e.g., Seth et al., U.S. Pat. No. 8,278,426), 4′-C(R_(a)R_(b))—N(R)—O-2′, 4′-C(R_(a)R_(b))—O—N(R)-2′, 4′-CH₂—O—N(R)-2′, and 4′-CH₂—N(R)—O-2′, wherein each R, R_(a), and R_(b) is, independently, H, a protecting group, or C₁-C₁₂ alkyl (see, e.g. Imanishi et al., U.S. Pat. No. 7,427,672).

In certain embodiments, such 4′ to 2′ bridges independently comprise from 1 to 4 linked groups independently selected from: —[C(R_(a))(R_(b))_(n)—, 4C(R_(a))(R_(b))]_(n)—O—, —C(R_(a))═C(R_(b))—, —C(R_(a))═N—, —C(═NR_(a))—, —C(═O)—, —C(═S)—, —O—, —Si(R_(a))₂—, —S(═O)_(x)—, and —N(R_(a))—;

wherein:

x is 0, 1, or 2;

n is 1, 2, 3, or 4;

each R_(a) and R_(b) is, independently, H, a protecting group, hydroxyl, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical, substituted C₅-C₇ alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), or sulfoxyl (S(═O)-J₁); and each J₁ and J₂ is, independently, H, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C₁-C₁₂ aminoalkyl, substituted C₁-C₁₂ aminoalkyl, or a protecting group.

Additional bicyclic sugar moieties are known in the art, see, for example: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. Acad. Sci. U.S.A., 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 2007, 129, 8362-8379; Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2, 558-561; Braasch et al., Chem. Biol., 2001, 8, 1-7; Orum et al., Curr. Opinion Mol. Ther., 2001, 3, 239-243; Wengel et al., U.S. Pat. No. 7,053,207, Imanishi et al., U.S. Pat. No. 6,268,490, Imanishi et al. U.S. Pat. No. 6,770,748, Imanishi et al., U.S. RE44,779; Wengel et al., U.S. Pat. No. 6,794,499, Wengel et al., U.S. Pat. No. 6,670,461; Wengel et al., U.S. Pat. No. 7,034,133, Wengel et al., U.S. Pat. No. 8,080,644; Wengel et al., U.S. Pat. No. 8,034,909; Wengel et al., U.S. Pat. No. 8,153,365; Wengel et al., U.S. Pat. No. 7,572,582; and Ramasamy et al., U.S. Pat. No. 6,525,191, Torsten et al., WO 2004/106356, Wengel et al., WO 1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. Pat. No. 7,547,684; Seth et al., U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat. No. 8,088,746; Seth et al., U.S. Pat. No. 7,750,131; Seth et al., U.S. Pat. No. 8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al., U.S. Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa et al., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805; Allerson et al., US2008/0039618; and Migawa et al., US2015/0191727.

In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, an LNA nucleoside (described herein) may be in the α-L configuration or in the β-D configuration.

α-L-methyleneoxy (4′-CH₂—O-2′) or α-L-LNA bicyclic nucleosides have been incorporated into oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). Herein, general descriptions of bicyclic nucleosides include both isomeric configurations. When the positions of specific bicyclic nucleosides (e.g., LNA or cEt) are identified in exemplified embodiments herein, they are in the β-D configuration, unless otherwise specified.

In certain embodiments, modified sugar moieties comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (e.g., 5′-substituted and 4′-2′ bridged sugars).

In certain embodiments, modified sugar moieties are sugar surrogates. In certain such embodiments, the oxygen atom of the sugar moiety is replaced, e.g., with a sulfur, carbon or nitrogen atom. In certain such embodiments, such modified sugar moieties also comprise bridging and/or non-bridging substituents as described herein. For example, certain sugar surrogates comprise a 4′-sulfur atom and a substitution at the 2′-position (see, e.g., Bhat et al., U.S. Pat. No. 7,875,733 and Bhat et al., U.S. Pat. No. 7,939,677) and/or the 5′ position.

In certain embodiments, sugar surrogates comprise rings having other than 5 atoms. For example, in certain embodiments, a sugar surrogate comprises a six-membered tetrahydropyran (“THP”). Such tetrahydropyrans may be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include but are not limited to hexitol nucleic acid (“HNA”), anitol nucleic acid (“ANA”), manitol nucleic acid (“MNA”) (see e.g., Leumann, C J. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoro HNA:

(“F-HNA”, see e.g., Swayze et al., U.S. Pat. No. 8,088,904; Swayze et al., U.S. Pat. No. 8,440,803; and Swayze et al., U.S. Pat. No. 9,005,906, F-HNA can also be referred to as a F-THP or 3′-fluoro tetrahydropyran), and nucleosides comprising additional modified THP compounds having the formula:

wherein, independently, for each of said modified THP nucleoside:

Bx is a nucleobase moiety;

T₃ and T₄ are each, independently, an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide or one of T₃ and T₄ is an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide and the other of T₃ and T₄ is H, a hydroxyl protecting group, a linked conjugate group, or a 5′ or 3′-terminal group; q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each, independently, H, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆ alkynyl, or substituted C₂-C₆ alkynyl; and each of R₁ and R₂ is independently selected from among: hydrogen, halogen, substituted or unsubstituted alkoxy, NJ₁J₂, SJ₁, N₃, OC(═X)J₁, OC(═X)NJ₁J₂, NJ₃C(═X)NJ₁J₂, and CN, wherein X is O, S or NJ₁, and each J₁, J₂, and J₃ is, independently, H or C₁-C₆ alkyl.

In certain embodiments, modified THP nucleosides are provided wherein q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and q₇ is other than H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and q₇ is methyl. In certain embodiments, modified THP nucleosides are provided wherein one of R₁ and R₂ is F. In certain embodiments, R₁ is F and R₂ is H, in certain embodiments, R₁ is methoxy and R₂ is H, and in certain embodiments, R₁ is methoxyethoxy and R₂ is H.

In certain embodiments, sugar surrogates comprise rings having more than 5 atoms and more than one heteroatom. For example, nucleosides comprising morpholino sugar moieties and their use in oligonucleotides have been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510 and Summerton et al., U.S. Pat. No. 5,698,685; Summerton et al., U.S. Pat. No. 5,166,315; Summerton et al., U.S. Pat. No. 5,185,444; and Summerton et al., U.S. Pat. No. 5,034,506). As used here, the term “morpholino” means a sugar surrogate having the following structure:

In certain embodiments, morpholinos may be modified, for example by adding or altering various substituent groups from the above morpholino structure. Such sugar surrogates are referred to herein as “modified morpholinos.”

In certain embodiments, sugar surrogates comprise acyclic moieties. Examples of nucleosides and oligonucleotides comprising such acyclic sugar surrogates include but are not limited to: peptide nucleic acid (“PNA”), acyclic butyl nucleic acid (see, e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoharan et al., US2013/130378.

Many other bicyclic and tricyclic sugar and sugar surrogate ring systems are known in the art that can be used in modified nucleosides.

2. Modified Nucleobases

Nucleobase (or base) modifications or substitutions are structurally distinguishable from, yet functionally interchangeable with, naturally occurring or synthetic unmodified nucleobases. Both natural and modified nucleobases are capable of participating in hydrogen bonding. Such nucleobase modifications can impart nuclease stability, binding affinity or some other beneficial biological property to antisense compounds.

In certain embodiments, compounds described herein comprise modified oligonucleotides. In certain embodiments, modified oligonucleotides comprise one or more nucleoside comprising an unmodified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleoside comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleoside that does not comprise a nucleobase, referred to as an abasic nucleoside.

In certain embodiments, modified nucleobases are selected from: 5-substituted pyrimidines, 6-azapyrimi┐dines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 2-aminopropyladenine, 5-hydroxymethyl cytosine, 5-methylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (C≡C—CH₃) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine-2-one, 1,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobases include those disclosed in Merigan et al., U.S. Pat. No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S. T., Ed., CRC Press, 2008, 163-166 and 442-443.

Publications that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include without limitation, Manoharan et al., US2003/0158403, Manoharan et al., US2003/0175906; Dinh et al., U.S. Pat. No. 4,845,205; Spielvogel et al., U.S. Pat. No. 5,130,302; Rogers et al., U.S. Pat. No. 5,134,066; Bischofberger et al., U.S. Pat. No. 5,175,273; Urdea et al., U.S. Pat. No. 5,367,066; Benner et al., U.S. Pat. No. 5,432,272; Matteucci et al., U.S. Pat. No. 5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cook et al., U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No. 5,484,908; Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al., U.S. Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540; Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat. No. 5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et al., U.S. Pat. No. 5,614,617; Froehler et al., U.S. Pat. No. 5,645,985; Cook et al., U.S. Pat. No. 5,681,941; Cook et al., U.S. Pat. No. 5,811,534; Cook et al., U.S. Pat. No. 5,750,692; Cook et al., U.S. Pat. No. 5,948,903; Cook et al., U.S. Pat. No. 5,587,470; Cook et al., U.S. Pat. No. 5,457,191; Matteucci et al., U.S. Pat. No. 5,763,588; Froehler et al., U.S. Pat. No. 5,830,653; Cook et al., U.S. Pat. No. 5,808,027; Cook et al., U.S. Pat. No. 6,166,199; and Matteucci et al., U.S. Pat. No. 6,005,096.

In certain embodiments, compounds targeted to a IRF4 nucleic acid comprise one or more modified nucleobases. In certain embodiments, the modified nucleobase is 5-methylcytosine. In certain embodiments, each cytosine is a 5-methylcytosine.

3. Modified Internucleoside Linkages

The naturally occurring internucleoside linkage of RNA and DNA is a 3′ to 5′ phosphodiester linkage In certain embodiments, compounds described herein having one or more modified, i.e. non-naturally occurring, internucleoside linkages are often selected over compounds having naturally occurring internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

Representative internucleoside linkages having a chiral center include but are not limited to alkylphosphonates and phosphorothioates. Modified oligonucleotides comprising internucleoside linkages having a chiral center can be prepared as populations of modified oligonucleotides comprising stereorandom internucleoside linkages, or as populations of modified oligonucleotides comprising phosphorothioate linkages in particular stereochemical configurations. In certain embodiments, populations of modified oligonucleotides comprise phosphorothioate internucleoside linkages wherein all of the phosphorothioate internucleoside linkages are stereorandom. Such modified oligonucleotides can be generated using synthetic methods that result in random selection of the stereochemical configuration of each phosphorothioate linkage. Nonetheless, as is well understood by those of skill in the art, each individual phosphorothioate of each individual oligonucleotide molecule has a defined stereoconfiguration. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising one or more particular phosphorothioate internucleoside linkages in a particular, independently selected stereochemical configuration. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 65% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 70% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 80% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 90% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 99% of the molecules in the population. Such chirally enriched populations of modified oligonucleotides can be generated using synthetic methods known in the art, e.g., methods described in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014), and WO 2017/015555. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one indicated phosphorothioate in the (Sp) configuration. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one phosphorothioate in the (Rp) configuration. In certain embodiments, modified oligonucleotides comprising (Rp) and/or (Sp) phosphorothioates comprise one or more of the following formulas, respectively, wherein “B” indicates a nucleobase:

Unless otherwise indicated, chiral internucleoside linkages of modified oligonucleotides described herein can be stereorandom or in a particular stereochemical configuration.

In certain embodiments, compounds targeted to an IRF4 nucleic acid comprise one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkages are phosphorothioate linkages. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

In certain embodiments, compounds described herein comprise oligonucleotides. Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom as well as internucleoside linkages that do not have a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known.

In certain embodiments, nucleosides of modified oligonucleotides may be linked together using any internucleoside linkage. The two main classes of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include but are not limited to phosphates, which contain a phosphodiester bond (“P═O”) (also referred to as unmodified or naturally occurring linkages), phosphotriesters, methylphosphonates, phosphoramidates, and phosphorothioates (“P═S”), and phosphorodithioates (“HS—P═S”). Representative non-phosphorus containing internucleoside linking groups include but are not limited to methylenemethylimino (—CH2-N(CH3)-O—CH2-), thiodiester, thionocarbamate (—O—C(═O)(NH)—S—); siloxane (—O—SiH2-O—); and N,N′-dimethylhydrazine (—CH2-N(CH3)-N(CH3)-). Modified internucleoside linkages, compared to naturally occurring phosphate linkages, can be used to alter, typically increase, nuclease resistance of the oligonucleotide. In certain embodiments, internucleoside linkages having a chiral atom can be prepared as a racemic mixture, or as separate enantiomers. Representative chiral internucleoside linkages include but are not limited to alkylphosphonates and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing internucleoside linkages are well known to those skilled in the art.

Neutral internucleoside linkages include, without limitation, phosphotriesters, methylphosphonates, MMI (3′-CH2-N(CH3)-O-5′), amide-3 (3′-CH2-C(═O)—N(H)-5′), amide-4 (3′-CH2-N(H)—C(═O)-5′), formacetal (3′-O—CH2-O-5), methoxypropyl, and thioformacetal (3′-S—CH2-O-5′). Further neutral internucleoside linkages include nonionic linkages comprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research; Y. S. Sanghvi and P. D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral internucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts.

In certain embodiments, oligonucleotides comprise modified internucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or modified internucleoside linkage motif. In certain embodiments, internucleoside linkages are arranged in a gapped motif. In such embodiments, the internucleoside linkages in each of two wing regions are different from the internucleoside linkages in the gap region. In certain embodiments the internucleoside linkages in the wings are phosphodiester and the internucleoside linkages in the gap are phosphorothioate. The nucleoside motif is independently selected, so such oligonucleotides having a gapped internucleoside linkage motif may or may not have a gapped nucleoside motif and if it does have a gapped nucleoside motif, the wing and gap lengths may or may not be the same.

In certain embodiments, oligonucleotides comprise a region having an alternating internucleoside linkage motif. In certain embodiments, oligonucleotides comprise a region of uniformly modified internucleoside linkages. In certain such embodiments, the oligonucleotide comprises a region that is uniformly linked by phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide is uniformly linked by phosphorothioate. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate and at least one internucleoside linkage is phosphorothioate.

In certain embodiments, the oligonucleotide comprises at least 6 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 8 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 10 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 6 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 8 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 10 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least block of at least one 12 consecutive phosphorothioate internucleoside linkages. In certain such embodiments, at least one such block is located at the 3′ end of the oligonucleotide. In certain such embodiments, at least one such block is located within 3 nucleosides of the 3′ end of the oligonucleotide.

In certain embodiments, oligonucleotides comprise one or more methylphosponate linkages. In certain embodiments, oligonucleotides having a gapmer nucleoside motif comprise a linkage motif comprising all phosphorothioate linkages except for one or two methylphosponate linkages. In certain embodiments, one methylphosponate linkage is in the central gap of an oligonucleotide having a gapmer nucleoside motif.

In certain embodiments, it is desirable to arrange the number of phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, it is desirable to arrange the number and position of phosphorothioate internucleoside linkages and the number and position of phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, the number of phosphorothioate internucleoside linkages may be decreased and the number of phosphodiester internucleoside linkages may be increased. In certain embodiments, the number of phosphorothioate internucleoside linkages may be decreased and the number of phosphodiester internucleoside linkages may be increased while still maintaining nuclease resistance. In certain embodiments it is desirable to decrease the number of phosphorothioate internucleoside linkages while retaining nuclease resistance. In certain embodiments it is desirable to increase the number of phosphodiester internucleoside linkages while retaining nuclease resistance.

Certain Motifs

In certain embodiments, compounds described herein comprise oligonucleotides. Oligonucleotides can have a motif, e.g. a pattern of unmodified and/or modified sugar moieties, nucleobases, and/or internucleoside linkages. In certain embodiments, modified oligonucleotides comprise one or more modified nucleoside comprising a modified sugar. In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more modified internucleoside linkage. In such embodiments, the modified, unmodified, and differently modified sugar moieties, nucleobases, and/or internucleoside linkages of a modified oligonucleotide define a pattern or motif. In certain embodiments, the patterns of sugar moieties, nucleobases, and internucleoside linkages are each independent of one another. Thus, a modified oligonucleotide may be described by its sugar motif, nucleobase motif and/or internucleoside linkage motif (as used herein, nucleobase motif describes the modifications to the nucleobases independent of the sequence of nucleobases).

a. Certain Sugar Motifs

In certain embodiments, compounds described herein comprise oligonucleotides. In certain embodiments, oligonucleotides comprise one or more type of modified sugar and/or unmodified sugar moiety arranged along the oligonucleotide or region thereof in a defined pattern or sugar motif. In certain instances, such sugar motifs include but are not limited to any of the sugar modifications discussed herein.

In certain embodiments, modified oligonucleotides comprise or consist of a region having a gapmer motif, which comprises two external regions or “wings” and a central or internal region or “gap.” The three regions of a gapmer motif (the 5′-wing, the gap, and the 3′-wing) form a contiguous sequence of nucleosides wherein at least some of the sugar moieties of the nucleosides of each of the wings differ from at least some of the sugar moieties of the nucleosides of the gap. Specifically, at least the sugar moieties of the nucleosides of each wing that are closest to the gap (the 3′-most nucleoside of the 5′-wing and the 5′-most nucleoside of the 3′-wing) differ from the sugar moiety of the neighboring gap nucleosides, thus defining the boundary between the wings and the gap (i.e., the wing/gap junction). In certain embodiments, the sugar moieties within the gap are the same as one another. In certain embodiments, the gap includes one or more nucleoside having a sugar moiety that differs from the sugar moiety of one or more other nucleosides of the gap. In certain embodiments, the sugar motifs of the two wings are the same as one another (symmetric gapmer). In certain embodiments, the sugar motif of the 5′-wing differs from the sugar motif of the 3′-wing (asymmetric gapmer).

In certain embodiments, the wings of a gapmer comprise 1-5 nucleosides. In certain embodiments, the wings of a gapmer comprise 2-5 nucleosides. In certain embodiments, the wings of a gapmer comprise 3-5 nucleosides. In certain embodiments, the nucleosides of a gapmer are all modified nucleosides.

In certain embodiments, the gap of a gapmer comprises 7-12 nucleosides. In certain embodiments, the gap of a gapmer comprises 7-10 nucleosides. In certain embodiments, the gap of a gapmer comprises 8-10 nucleosides. In certain embodiments, the gap of a gapmer comprises 10 nucleosides. In certain embodiment, each nucleoside of the gap of a gapmer is an unmodified 2′-deoxy nucleoside.

In certain embodiments, the gapmer is a deoxy gapmer. In such embodiments, the nucleosides on the gap side of each wing/gap junction are unmodified 2′-deoxy nucleosides and the nucleosides on the wing sides of each wing/gap junction are modified nucleosides. In certain such embodiments, each nucleoside of the gap is an unmodified 2′-deoxy nucleoside. In certain such embodiments, each nucleoside of each wing is a modified nucleoside.

In certain embodiments, a modified oligonucleotide has a fully modified sugar motif wherein each nucleoside of the modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a region having a fully modified sugar motif wherein each nucleoside of the region comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a region having a fully modified sugar motif, wherein each nucleoside within the fully modified region comprises the same modified sugar moiety, referred to herein as a uniformly modified sugar motif. In certain embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In certain embodiments, each nucleoside of a uniformly modified comprises the same 2′-modification.

In certain embodiments, a modified oligonucleotide can comprise a sugar motif described in Swayze et al., US2010/0197762; Freier et al., US2014/0107330; Freier et al., US2015/0184153; and Seth et al., US2015/0267195, each of which is incorporated by reference in its entirety herein.

Certain embodiments provided herein are directed to modified oligomeric compounds useful for inhibiting target nucleic acid expression, which can be useful for treating, preventing, ameliorating, or slowing progression of a disease associated with such a target nucleic acid. In certain embodiments, the modified oligomeric compounds comprise antisense oligonucleotides that are gapmers having certain sugar motifs. In certain embodiments, the gapmer sugar motifs provided herein can be combined with any nucleobase sequence and any internucleoside linkage motif to form potent antisense oligonucleotides.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: ekk-d9-kkee, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: k-d9-kekeke, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: kkk-d8-kekek, wherein ‘d’ represents a 2′-deoxyribose sugar, 1′ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: kkk-d9-keke, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: kk-d9-kdkdk, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a compound comprises a modified oligonucleotide 16 linked nucleosides in length having the motif: kk-d9-eeekk, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: kk-d9-eeekk, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

In certain embodiments, a method comprises contacting a cell or administering to a subject a compound comprising a modified oligonucleotide 16 linked nucleosides in length having the motif: kk-d9-ekeke, wherein ‘d’ represents a 2′-deoxyribose sugar, ‘k’ represents a cEt nucleoside, and ‘e’ represents a 2′-MOE nucleoside. In certain embodiments, the cell is a cancer cell. In certain embodiments, the subject has cancer. In certain embodiments, administering the compound to the subject treats the subject's cancer.

b. Certain Nucleobase Motifs

In certain embodiments, compounds described herein comprise oligonucleotides. In certain embodiments, oligonucleotides comprise modified and/or unmodified nucleobases arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases are modified. In certain embodiments, each purine or each pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleobases in a modified oligonucleotide are 5-methylcytosines.

In certain embodiments, modified oligonucleotides comprise a block of modified nucleobases. In certain such embodiments, the block is at the 3′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 3′-end of the oligonucleotide. In certain embodiments, the block is at the 5′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 5′-end of the oligonucleotide.

In certain embodiments, oligonucleotides having a gapmer motif comprise a nucleoside comprising a modified nucleobase. In certain such embodiments, one nucleoside comprising a modified nucleobase is in the central gap of an oligonucleotide having a gapmer motif. In certain such embodiments, the sugar moiety of said nucleoside is a 2′-deoxyribosyl moiety. In certain embodiments, the modified nucleobase is selected from: a 2-thiopyrimidine and a 5-propynepyrimidine.

c. Certain Internucleoside Linkage Motifs

In certain embodiments, compounds described herein comprise oligonucleotides. In certain embodiments, oligonucleotides comprise modified and/or unmodified internucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, essentially each internucleoside linking group is a phosphate internucleoside linkage (P═O). In certain embodiments, each internucleoside linking group of a modified oligonucleotide is a phosphorothioate (P═S). In certain embodiments, each internucleoside linking group of a modified oligonucleotide is independently selected from a phosphorothioate and phosphate internucleoside linkage. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer and the internucleoside linkages within the gap are all modified. In certain such embodiments, some or all of the internucleoside linkages in the wings are unmodified phosphate linkages. In certain embodiments, the terminal internucleoside linkages are modified.

4. Certain Modified Oligonucleotides

In certain embodiments, compounds described herein comprise modified oligonucleotides. In certain embodiments, the above modifications (sugar, nucleobase, internucleoside linkage) are incorporated into a modified oligonucleotide. In certain embodiments, modified oligonucleotides are characterized by their modification, motifs, and overall lengths. In certain embodiments, such parameters are each independent of one another. Thus, unless otherwise indicated, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif may be modified or unmodified and may or may not follow the gapmer modification pattern of the sugar modifications. For example, the internucleoside linkages within the wing regions of a sugar gapmer may be the same or different from one another and may be the same or different from the internucleoside linkages of the gap region of the sugar motif. Likewise, such gapmer oligonucleotides may comprise one or more modified nucleobase independent of the gapmer pattern of the sugar modifications. Furthermore, in certain instances, an oligonucleotide is described by an overall length or range and by lengths or length ranges of two or more regions (e.g., a regions of nucleosides having specified sugar modifications), in such circumstances it may be possible to select numbers for each range that result in an oligonucleotide having an overall length falling outside the specified range. In such circumstances, both elements must be satisfied. For example, in certain embodiments, a modified oligonucleotide consists of 15-20 linked nucleosides and has a sugar motif consisting of three regions, A, B, and C, wherein region A consists of 2-6 linked nucleosides having a specified sugar motif, region B consists of 6-10 linked nucleosides having a specified sugar motif, and region C consists of 2-6 linked nucleosides having a specified sugar motif. Such embodiments do not include modified oligonucleotides where A and C each consist of 6 linked nucleosides and B consists of 10 linked nucleosides (even though those numbers of nucleosides are permitted within the requirements for A, B, and C) because the overall length of such oligonucleotide is 22, which exceeds the upper limit of the overall length of the modified oligonucleotide (20). Herein, if a description of an oligonucleotide is silent with respect to one or more parameter, such parameter is not limited. Thus, a modified oligonucleotide described only as having a gapmer sugar motif without further description may have any length, internucleoside linkage motif, and nucleobase motif. Unless otherwise indicated, all modifications are independent of nucleobase sequence.

Certain Conjugated Compounds

In certain embodiments, the compounds described herein comprise or consist of an oligonucleotide (modified or unmodified) and optionally one or more conjugate groups and/or terminal groups. Conjugate groups consist of one or more conjugate moiety and a conjugate linker which links the conjugate moiety to the oligonucleotide. Conjugate groups may be attached to either or both ends of an oligonucleotide and/or at any internal position. In certain embodiments, conjugate groups are attached to the 2′-position of a nucleoside of a modified oligonucleotide. In certain embodiments, conjugate groups that are attached to either or both ends of an oligonucleotide are terminal groups. In certain such embodiments, conjugate groups or terminal groups are attached at the 3′ and/or 5′-end of oligonucleotides. In certain such embodiments, conjugate groups (or terminal groups) are attached at the 3′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 3′-end of oligonucleotides. In certain embodiments, conjugate groups (or terminal groups) are attached at the 5′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 5′-end of oligonucleotides.

In certain embodiments, the oligonucleotide is modified. In certain embodiments, the oligonucleotide of a compound has a nucleobase sequence that is complementary to a target nucleic acid. In certain embodiments, oligonucleotides are complementary to a messenger RNA (mRNA). In certain embodiments, oligonucleotides are complementary to a sense transcript.

Examples of terminal groups include but are not limited to conjugate groups, capping groups, phosphate moieties, protecting groups, modified or unmodified nucleosides, and two or more nucleosides that are independently modified or unmodified.

A. Certain Conjugate Groups

In certain embodiments, oligonucleotides are covalently attached to one or more conjugate groups. In certain embodiments, conjugate groups modify one or more properties of the attached oligonucleotide, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge and clearance In certain embodiments, conjugate groups impart a new property on the attached oligonucleotide, e.g., fluorophores or reporter groups that enable detection of the oligonucleotide.

Certain conjugate groups and conjugate moieties have been described previously, for example: cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., do-decan-diol or undecyl residues (Saison-Behmoaras et al., EMBO 1, 1991, 10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic, a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, i, 923-937),_a tocopherol group (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4, e220; doi:10.1038/mtna.2014.72 and Nishina et al., Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (e.g., WO2014/179620).

1. Conjugate Moieties

Conjugate moieties include, without limitation, intercalators, reporter molecules, polyamines, polyamides, peptides, carbohydrates (e.g., GalNAc), vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.

In certain embodiments, a conjugate moiety comprises an active drug substance, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic acid, a benzothiadiazide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.

2. Conjugate Linkers

Conjugate moieties are attached to oligonucleotides through conjugate linkers. In certain compounds, a conjugate group is a single chemical bond (i.e. conjugate moiety is attached to an oligonucleotide via a conjugate linker through a single bond). In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units.

In certain embodiments, a conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group. In certain embodiments, the conjugate linker includes at least one neutral linking group.

In certain embodiments, conjugate linkers, including the conjugate linkers described above, are bifunctional linking moieties, e.g., those known in the art to be useful for attaching conjugate groups to parent compounds, such as the oligonucleotides provided herein. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a particular site on a compound and the other is selected to bind to a conjugate group. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.

Examples of conjugate linkers include but are not limited to pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include but are not limited to substituted or unsubstituted C₁-C₁₀ alkyl, substituted or unsubstituted C₂-C₁₀ alkenyl or substituted or unsubstituted C₂-C₁₀ alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

In certain embodiments, conjugate linkers comprise 1-10 linker-nucleosides. In certain embodiments, such linker-nucleosides are modified nucleosides. In certain embodiments such linker-nucleosides comprise a modified sugar moiety. In certain embodiments, linker-nucleosides are unmodified. In certain embodiments, linker-nucleosides comprise an optionally protected heterocyclic base selected from a purine, substituted purine, pyrimidine or substituted pyrimidine. In certain embodiments, a cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typically desirable for linker-nucleosides to be cleaved from the compound after it reaches a target tissue. Accordingly, linker-nucleosides are typically linked to one another and to the remainder of the compound through cleavable bonds. In certain embodiments, such cleavable bonds are phosphodiester bonds.

Herein, linker-nucleosides are not considered to be part of the oligonucleotide. Accordingly, in embodiments in which a compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and/or a specified percent complementarity to a reference nucleic acid and the compound also comprises a conjugate group comprising a conjugate linker comprising linker-nucleosides, those linker-nucleosides are not counted toward the length of the oligonucleotide and are not used in determining the percent complementarity of the oligonucleotide for the reference nucleic acid. For example, a compound may comprise (1) a modified oligonucleotide consisting of 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker-nucleosides that are contiguous with the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such a compound is more than 30. Alternatively, an compound may comprise a modified oligonucleotide consisting of 8-30 nucleosides and no conjugate group. The total number of contiguous linked nucleosides in such a compound is no more than 30. Unless otherwise indicated conjugate linkers comprise no more than 10 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 5 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 2 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 1 linker-nucleoside.

In certain embodiments, it is desirable for a conjugate group to be cleaved from the oligonucleotide. For example, in certain circumstances compounds comprising a particular conjugate moiety are better taken up by a particular cell type, but once the compound has been taken up, it is desirable that the conjugate group be cleaved to release the unconjugated or parent oligonucleotide. Thus, certain conjugate may comprise one or more cleavable moieties, typically within the conjugate linker. In certain embodiments, a cleavable moiety is a cleavable bond. In certain embodiments, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain embodiments, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.

In certain embodiments, a cleavable bond is selected from among: an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide. In certain embodiments, a cleavable bond is one or both of the esters of a phosphodiester. In certain embodiments, a cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a phosphate linkage between an oligonucleotide and a conjugate moiety or conjugate group.

In certain embodiments, a cleavable moiety comprises or consists of one or more linker-nucleosides. In certain such embodiments, one or more linker-nucleosides are linked to one another and/or to the remainder of the compound through cleavable bonds. In certain embodiments, such cleavable bonds are unmodified phosphodiester bonds. In certain embodiments, a cleavable moiety is 2′-deoxy nucleoside that is attached to either the 3′ or 5′-terminal nucleoside of an oligonucleotide by a phosphate internucleoside linkage and covalently attached to the remainder of the conjugate linker or conjugate moiety by a phosphate or phosphorothioate linkage. In certain such embodiments, the cleavable moiety is 2′-deoxyadenosine.

Compositions and Methods for Formulating Pharmaceutical Compositions

Compounds described herein may be admixed with pharmaceutically acceptable active or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

Certain embodiments provide pharmaceutical compositions comprising one or more compounds or a salt thereof. In certain embodiments, the compounds are antisense compounds or oligomeric compounds. In certain embodiments, the compounds comprise or consist of a modified oligonucleotide. In certain such embodiments, the pharmaceutical composition comprises a suitable pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition comprises a sterile saline solution and one or more compound. In certain embodiments, such pharmaceutical composition consists of a sterile saline solution and one or more compound. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises one or more compound and sterile water. In certain embodiments, a pharmaceutical composition consists of one compound and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises one or more compound and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more compound and sterile PBS. In certain embodiments, the sterile PBS is pharmaceutical grade PBS. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

A compound described herein targeted to IRF4 nucleic acid can be utilized in pharmaceutical compositions by combining the compound with a suitable pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutically acceptable diluent is water, such as sterile water suitable for injection. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising a compound targeted to IRF4 nucleic acid and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is water. In certain embodiments, the compound comprises or consists of a modified oligonucleotide provided herein.

Pharmaceutical compositions comprising compounds provided herein encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. In certain embodiments, the compounds are antisense compounds or oligomeric compounds. In certain embodiments, the compound comprises or consists of a modified oligonucleotide. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

A prodrug can include the incorporation of additional nucleosides at one or both ends of a compound which are cleaved by endogenous nucleases within the body, to form the active compound. In certain embodiments, the compounds or compositions further comprise a pharmaceutically acceptable carrier or diluent.

EXAMPLES

The Examples below describe the screening process to identify lead compounds targeted to IRF4. Out of over 3,000 oligonucleotides that were screened, ION 690890, 935658, 935696, 935762, 935918, 935968, 882800, 1012795, 1014095, and 1014834 emerged as the top lead compounds. In particular, ION 935918 exhibited the best combination of properties in terms of potency and tolerability out of over 3,000 oligonucleotides.

Non-Limiting Disclosure and Incorporation by Reference

Although the sequence listing accompanying this filing identifies each sequence as either “RNA” or “DNA” as required, in reality, those sequences may be modified with any combination of chemical modifications. One of skill in the art will readily appreciate that such designation as “RNA” or “DNA” to describe modified oligonucleotides is, in certain instances, arbitrary. For example, an oligonucleotide comprising a nucleoside comprising a 2′-OH sugar moiety and a thymine base could be described as a DNA having a modified sugar (2′-OH for the natural 2′-H of DNA) or as an RNA having a modified base (thymine (methylated uracil) for natural uracil of RNA).

Accordingly, nucleic acid sequences provided herein, including, but not limited to those in the sequence listing, are intended to encompass nucleic acids containing any combination of natural or modified RNA and/or DNA, including, but not limited to such nucleic acids having modified nucleobases. By way of further example and without limitation, an oligonucleotide having the nucleobase sequence “ATCGATCG” encompasses any oligonucleotides having such nucleobase sequence, whether modified or unmodified, including, but not limited to, such compounds comprising RNA bases, such as those having sequence “AUCGAUCG” and those having some DNA bases and some RNA bases such as “AUCGATCG” and compounds having other modified nucleobases, such as “AT^(m)CGAUCG,” wherein ^(m)C indicates a cytosine base comprising a methyl group at the 5-position.

While certain compounds, compositions and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references recited in the present application is incorporated herein by reference in its entirety.

Example 1: Effect of 5-10-5 MOE Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 60,000 cells per well were transfected using electroporation with 20,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (forward sequence AAGCCTTGGCGTTCTCAGACT, designated herein as SEQ ID NO: 3386; reverse sequence TCAGCTCCTTCACGAGGATTTC, designated herein as SEQ ID NO: 3387; probe sequence CCGGCTGCACATCTGCCTGTACTACC, designated herein as SEQ ID: 3388) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Table 1 are 5-10-5 MOE gapmers. The gapmers are 20 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising five 2′-MOE nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): eeeeeddddddddddeeeee; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘e’ represents a 2′-MOE modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Table 1 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 1 Percent control of human IRF4 mRNA with 5-10-5 MOE gapmers with phosphorothioate internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID Compound NO: 1 NO: 1 NO: 2 NO: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 434887  164  183  5191  5210 GCAGCTCACCGCGCTCATGC 57  3 434888  175  194  5202  5221 TTCCCGTTGCCGCAGCTCAC 34  4 434889  190  209  5217  5236 AGCCACTGGCGGAGCTTCCC 43  5 434890  294  313  5321  5340 TGTAGTCCTGCTTGCCCGCG 51  6 434891  304  323  5331  5350 TCCTCGCGGTTGTAGTCCTG 53  7 434892  330  349 N/A N/A CCCAAGCCTTGAAGAGCGCG 44  8 434893  367  386  6846  6865 TTGTCGATGCCTTCTCGGAA 47  9 434894  375  394  6854  6873 GGTCCGGCTTGTCGATGCCT 40 10 434895  430  449  6909  6928 TCAAAGTCATTGCTCTTGTT 55 11 434896  436  455  6915  6934 AGTTCCTCAAAGTCATTGCT 42 12 434897  442  461  6921  6940 TCAACCAGTTCCTCAAAGTC 75 13 434898  447  466  6926  6945 TCCGCTCAACCAGTTCCTCA 32 14 434899  474  493  6953  6972 TGTACGGGTCTGAGATGTCC 44 15 434900  532  551  7850  7869 TCCAGGGTGAGCTGCTTGGC 43 16 434901  555  574  7873  7892 GGCTCATGGACATCTGCGGG 45 17 434902  675  694  9165  9184 TTTCCGGGTGTGGCTGATCC 36 18 434903  690  709  9180  9199 GACATTGGTACGGGATTTCC 38 19 434904  732  751  9222  9241 CTGGGCCTTGCCAGTGGTGG 59 20 434905  739  758  9229  9248 TCACAAGCTGGGCCTTGCCA 30 21 434906  744  763  9234  9253 CATTTTCACAAGCTGGGCCT 37 22 434907  751  770 N/A N/A TGGCAACCATTTTCACAAGC 37 23 434908  758  777 N/A N/A TGTCACCTGGCAACCATTTT 49 24 434909  778  797 10843 10862 GCACAAGCATAAAAGGTTCC 26 25 434916  940  959 13493 13512 TGGGAGATCCGGCAGCCCTC 66 26 434917 1004 1023 13557 13576 GCCATTGTCCTCTGGGTAGG 38 27 434918 1042 1061 13595 13614 TCCAGGTGGCTCAGCAGCTT 25 28 434919 1063 1082 13616 13635 ATCCAGAGGACCACGCCCCT 78 29 434920 1068 1087 13621 13640 GGGCCATCCAGAGGACCACG 36 30 434921 1119 1138 13672 13691 CGTCCCAGTAGATCCTGCTC 46 31 434922 1187 1206 13740 13759 GTCAAAGAGCTTGCAGGTCT 32 32 434923 1192 1211 13745 13764 TGTGTGTCAAAGAGCTTGCA 17 33 434924 1344 1363 19461 19480 GTTGTCTGGCTAGCAGAGGT 33 34 434925 1364 1383 19481 19500 TTGTTGAGCAAAATAATATA 97 35 434926 1391 1410 19508 19527 GTAGCCCCTCAGGAAATGTC 15 36 434927 1420 1439 19537 19556 TCTGGATTGCTGATGTGTTC 46 37 434928 1430 1449 19547 19566 GTGGTAATCTTCTGGATTGC 42 38 434929 1450 1469 19567 19586 GAGGAATGGCGGATAGATCT 63 39 434930 1707 1726 19824 19843 CACTAAAGTCAAATATTTAC 88 40 434931 1712 1731 19829 19848 GCTTTCACTAAAGTCAAATA 29 41 434932 1821 1840 19938 19957 CAGATGTCACTGATTTTCCA 41 42 434933 1826 1845 19943 19962 CCAATCAGATGTCACTGATT 44 43 434934 1838 1857 19955 19974 TAAGCTCATCTGCCAATCAG 71 44 434935 2196 2215 20313 20332 CCAAGGCTACAGGCACGGCT 29 45 434936 2234 2253 20351 20370 ACACCAGGAAACCGCTGGCA 37 46 434937 2293 2312 20410 20429 CTTCCAGGAAAGGCCAAGGA 23 47 434938 2356 2375 20473 20492 TGTCCCATCCAAGAGTAGCG 30 48 434939 2662 2681 20779 20798 CTTCCAGTGGTGGGTCCTGG 53 49 434940 2705 2724 20822 20841 AACAGCCCACTGAGTGTGCA 43 50 434941 2711 2730 20828 20847 AGCAGAAACAGCCCACTGAG 58 51 434942 3435 3454 21552 21571 CTGGGTACATGGCAGTGGAG 27 52 434943 3805 3824 21922 21941 GCATTTTCCAGAAAATTCAG 33 53 434944 4477 4496 22594 22613 CCCAGAGTTGTTCCACCCCT 21 54 434945 N/A N/A 22826 22845 GCTGGCCACAGAGGACTTCG 21 55 434946 4737 4756 22854 22873 GCCTTCACGCACCATTCAGA 52 56 434947 4812 4831 22929 22948 CCACCTGCATCGAGATCAGT 31 57 434948 4818 4837 22935 22954 GGAGATCCACCTGCATCGAG 32 58 434949 5035 5054 23152 23171 GGAAGTGGACCCCATTGCCT 17 59 434952 N/A N/A 18834 18853 ATCTAAGGCAAGCTGAATGC 67 60 434953 N/A N/A 18839 18858 ACAGCATCTAAGGCAAGCTG 38 61 434954 N/A N/A 18845 18864 GAATTTACAGCATCTAAGGC 52 62 434955 N/A N/A 18850 18869 TTCCTGAATTTACAGCATCT 21 63 434957 N/A N/A  5359  5378 GCCGGAGACCTTGAAGAGCG 94 64 434958 N/A N/A  7480  7499 ACTGGTCAGAATCTTGAAAA 94 65 434959 N/A N/A  9101  9120 GTTGTGAACCTGCTAAAGGA 90 66 434960 N/A N/A 10818 10837 CCTGGCAACCTGCATTTGCA 43 67 434961 N/A N/A 10823 10842 TGTCACCTGGCAACCTGCAT 45 68 434962 N/A N/A 12030 12049 GGGCAGGTTTCATTTCATTT 33 69 434963 N/A N/A 13414 13433 GCCGGCAGTCTGCAAACACA 58 70 434964 N/A N/A 19448 19467 CAGAGGTTCTACCTTTAATA 50 71

Example 2: Effect of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 20,000 cells per well were transfected using electroporation with 4,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 2 and 3 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 2 and 3 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 2 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609281  156  171  5183  5198 GCTCATGCCGAACTCT  81  72 609282  213  228  5240  5255 GCTGTCGATCTGGTCG  91  73 609283  216  231  5243  5258 GCCGCTGTCGATCTGG  76  74 609284  219  234  5246  5261 CTTGCCGCTGTCGATC  75  75 609285  222  237  5249  5264 GTACTTGCCGCTGTCG  73  76 609286  238  253  5265  5280 CCCACACCAGCCCGGG 106  77 609287  241  256  5268  5283 TCTCCCACACCAGCCC  94  78 609288  244  259  5271  5286 CGTTCTCCCACACCAG  52  79 609289  294  309  5321  5336 GTCCTGCTTGCCCGCG  69  80 609290  297  312  5324  5339 GTAGTCCTGCTTGCCC  56  81 609291  334  349 N/A N/A CCCAAGCCTTGAAGAG  94  82 609292  431  446  6910  6925 AAGTCATTGCTCTTGT  72  83 609293  434  449  6913  6928 TCAAAGTCATTGCTCT  79  84 609294  437  452  6916  6931 TCCTCAAAGTCATTGC  68  85 609295  496  511  6975  6990 GAACAATCCTGTACAC 105  86 609296  514  529  6993  7008 CTTTTTTGGCTCCCTC  64  87 609297  517  532 N/A N/A CTCCTTTTTTGGCTCC  81  88 609298  610  625 N/A N/A GAACCTGCTGGGCTGG  64  89 609299  730  745  9220  9235 CTTGCCAGTGGTGGCC  84  90 609300  733  748  9223  9238 GGCCTTGCCAGTGGTG 112  91 609301  752  767 N/A N/A CAACCATTTTCACAAG  83  92 609302  755  770 N/A N/A TGGCAACCATTTTCAC  93  93 609303  758  773 N/A N/A ACCTGGCAACCATTTT  96  94 609304  761  776 N/A N/A GTCACCTGGCAACCAT  59  95 609305  764  779 10829 10844 CCTGTCACCTGGCAAC  59  96 609306  767  782 10832 10847 GTTCCTGTCACCTGGC  51  97 609307  770  785 10835 10850 AAGGTTCCTGTCACCT 102  98 609308  773  788 10838 10853 TAAAAGGTTCCTGTCA  94  99 609309  776  791 10841 10856 GCATAAAAGGTTCCTG  74 100 609310  780  795 10845 10860 ACAAGCATAAAAGGTT  96 101 609311  799  814 10864 10879 CCTGGGACTCAGGTGG  41 102 609312  802  817 10867 10882 GAGCCTGGGACTCAGG  51 103 609313  832  847 10897 10912 ACCTTATGCTTGGCTC  69 104 609314  835  850 10900 10915 CAGACCTTATGCTTGG  90 105 609317  943  958 13496 13511 GGGAGATCCGGCAGCC 110 106 609318  979  994 13532 13547 CCTGGTCCAGGTTGCT  74 107 609319  982  997 13535 13550 GGACCTGGTCCAGGTT 102 108 609320  985 1000 13538 13553 ACAGGACCTGGTCCAG  65 109 609321 1046 1061 13599 13614 TCCAGGTGGCTCAGCA  59 110 609322 1049 1064 13602 13617 CTCTCCAGGTGGCTCA  87 111 609323 1052 1067 13605 13620 CCCCTCTCCAGGTGGC  87 112 609324 1194 1209 13747 13762 TGTGTCAAAGAGCTTG  78 113 609325 1198 1213 13751 13766 GCTGTGTGTCAAAGAG  94 114 609326 1201 1216 13754 13769 ACTGCTGTGTGTCAAA 106 115 609327 1264 1279 17057 17072 GAGTCACCTGGAATCT  57 116 609328 1291 1306 17084 17099 GGTCTGGAAACTCCTC  48 117 609329 1294 1309 17087 17102 GAGGGTCTGGAAACTC 104 118 609330 1297 1312 17090 17105 TCTGAGGGTCTGGAAA  91 119 609331 1321 1336 17114 17129 GAGCTGTGATGAGCTT  90 120 609332 1342 1357 19459 19474 TGGCTAGCAGAGGTTC  44 121 609333 1345 1360 19462 19477 GTCTGGCTAGCAGAGG  34 122 609334 1348 1363 19465 19480 GTTGTCTGGCTAGCAG  41 123 609335 1389 1404 19506 19521 CCTCAGGAAATGTCCA  63 124 609336 1393 1408 19510 19525 AGCCCCTCAGGAAATG  76 125 609337 2068 2083 20185 20200 CTTCCCTGAGAAATGG  50 126 609338 2071 2086 20188 20203 TTACTTCCCTGAGAAA  80 127 609339 2719 2734 20836 20851 AATAAGCAGAAACAGC 101 128 609340 3533 3548 21650 21665 ATCTATAAGATGTATA 101 129 609341 3536 3551 21653 21668 TGCATCTATAAGATGT  68 130 609342 3803 3818 21920 21935 TCCAGAAAATTCAGCT  52 131 609343 3809 3824 21926 21941 GCATTTTCCAGAAAAT  36 132 609344 N/A N/A  6995  7010 ACCTTTTTTGGCTCCC  79 133 609345 N/A N/A  7740  7755 ATTCTTAGAATGCATA  62 134 609346 N/A N/A  8504  8519 CAAACTCCTCAGGGAA  77 135 609347 N/A N/A  9242  9257 TTACCATTTTCACAAG  92 136 609348 N/A N/A 10796 10811 AATCAGATGATGTCTA 103 137 609349 N/A N/A 10813 10828 CTGCATTTGCAAATAA 121 138 609350 N/A N/A 10819 10834 GGCAACCTGCATTTGC 113 139 609351 N/A N/A 10825 10840 TCACCTGGCAACCTGC  75 140 609352 N/A N/A 11951 11966 CACCCACACAAGTCTT  91 141 609353 N/A N/A 11972 11987 GTTTATTTCCTACTCT  84 142 609354 N/A N/A 11978 11993 ATAGCTGTTTATTTCC  38 143 609355 N/A N/A 11985 12000 GATATAAATAGCTGTT  63 144 609356 N/A N/A 12024 12039 CATTTCATTTAATGTC  67 145 609357 N/A N/A 12031 12046 CAGGTTTCATTTCATT  40 146 609358 N/A N/A 13795 13810 CCTCCATTCTAACAGA 118 147

TABLE 3 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609360 4711 4726 22828 22843 TGGCCACAGAGGACTT  67 148 609361    1   16  3740  3755 ACTGAGAGTGCGAGGT 120 149 609362  121  136  5148  5163 CCAGGTTCATGCCCCG  46 150 609363  181  196  5208  5223 GCTTCCCGTTGCCGCA  82 151 609364  300  315  5327  5342 GTTGTAGTCCTGCTTG  78 152 609365  420  435  6899  6914 CTTGTTCAAAGCGCAC  68 153 609366  480  495  6959  6974 TTTGTACGGGTCTGAG  80 154 609367  672  687  9162  9177 GTGTGGCTGATCCGGG  73 155 609368  736  751  9226  9241 CTGGGCCTTGCCAGTG 114 156 609369  810  825 10875 10890 GACTCCGGGAGCCTGG  35 157 609371 1004 1019 13557 13572 TTGTCCTCTGGGTAGG  70 158 609372 1124 1139 13677 13692 CCGTCCCAGTAGATCC  75 159 609373 1184 1199 13737 13752 AGCTTGCAGGTCTGGT  47 160 609374 1431 1446 19548 19563 GTAATCTTCTGGATTG  71 161 609375 1527 1542 19644 19659 TCCCCGTATCAAAAAA 112 162 609376 1682 1697 19799 19814 CACTTGTCTTGGGTGG  55 163 609377 1742 1757 19859 19874 AACAGTAAGAGGGCAG  37 164 609378 1866 1881 19983 19998 GCAAAGCCACCCTTCC  40 165 609379 1986 2001 20103 20118 TCTTCCAGCAAGACCT  72 166 609380 2111 2126 20228 20243 ATACATTTCTTTTACG  78 167 609381 2171 2186 20288 20303 GATTCATTTCCTTCAC  51 168 609382 2231 2246 20348 20363 GAAACCGCTGGCAGGT  50 169 609383 2295 2310 20412 20427 TCCAGGAAAGGCCAAG  51 170 609384 2304 2319 20421 20436 TAACTGGCTTCCAGGA  70 171 609385 2365 2380 20482 20497 AAAAATGTCCCATCCA  70 172 609386 2431 2446 20548 20563 GTCAAAAAGATGCAGA  50 173 609387 2494 2509 20611 20626 GATTTATGTTCCTTAA  53 174 609388 2574 2589 20691 20706 AACAAACAGAGGAGCG  84 175 609389 2634 2649 20751 20766 GCCTGGGAGTCCCCGG  82 176 609390 2694 2709 20811 20826 GTGCAGTTCCGTAGTC  79 177 609391 2754 2769 20871 20886 ACTATAATTGGCACGA  31 178 609392 2816 2831 20933 20948 TCTTTGGGATTCTATA  40 179 609393 2936 2951 21053 21068 GGAGTAATAGTAAATA  64 180 609394 3081 3096 21198 21213 CTGCTCACTAAGCTTG  27 181 609395 3147 3162 21264 21279 GTATTAATATTCTGAC  37 182 609396 3216 3231 21333 21348 GGAGATCCTTTTTATT  68 183 609397 3336 3351 21453 21468 GACTCCATGAGGTTTT  30 184 609398 3437 3452 21554 21569 GGGTACATGGCAGTGG  32 185 609399 3591 3606 21708 21723 GCAGTTCTTAATATCA  40 186 609400 3657 3672 21774 21789 TGAAGTGCTGTGTGGG  43 187 609401 3717 3732 21834 21849 TCCGCTTGGAGAATTA  87 188 609402 3782 3797 21899 21914 GTTAAAGCAGCATAAT  73 189 609403 3851 3866 21968 21983 AGATGTAAAGATAGGA  46 190 609404 3911 3926 22028 22043 AGTTCATTCCCTAGGT  56 191 609405 3971 3986 22088 22103 GTTCCTTTTCAGAGTC  28 192 609406 4031 4046 22148 22163 AGTACAAACTAAATTC  76 193 609407 4166 4181 22283 22298 GAGGTTTTCCTAAATA  31 194 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  21 195 609409 4359 4374 22476 22491 CTGTAGGATTTTACAT  76 196 609410 4479 4494 22596 22611 CAGAGTTGTTCCACCC  38 197 609411 4599 4614 22716 22731 AATGAACGGAAGTTTA  67 198 609412 4665 4680 22782 22797 AACCAATCCCAACACT  88 199 609413 4727 4742 22844 22859 TTCAGACAGATGCAGC  38 200 609414 4800 4815 22917 22932 CAGTCTCAAAAACGGG  49 201 609415 4862 4877 22979 22994 ACCTTTACTTCATTCC  36 202 609416 4987 5002 23104 23119 ACGGGAATTTCCATTG  33 203 609417 5037 5052 23154 23169 AAGTGGACCCCATTGC  69 204 609418 5077 5092 23194 23209 TTCAGCAGAAAGTGGG  52 205 609419 5146 5161 23263 23278 CGCAGAGCCAGTAGGG  33 206 609420 N/A N/A  2604  2619 AAAAGCCCAAAATAGG 112 207  2875  2890 609421 N/A N/A  8538  8553 CTGGCATTGAGACGGG  39 208  8746  8761  8850  8865  8902  8917  9058  9073 609422 N/A N/A  8542  8557 AGCACTGGCATTGAGA  26 209  8750  8765  8854  8869  8906  8921  9062  9077 609423 N/A N/A  8547  8562 TAAGAAGCACTGGCAT  74 210  8703  8718  8807  8822  8963  8978  9067  9082 609424 N/A N/A  8552  8567 TGAGATAAGAAGCACT  80 211  8708  8723  8812  8827  8968  8983  9072  9087 609425 N/A N/A  8560  8575 GGAGAGGCTGAGATAA  71 212  8716  8731  8820  8835  8976  8991  9080  9095 609426 N/A N/A  8561  8576 AGGAGAGGCTGAGATA  79 213  8613  8628  8665  8680  8717  8732  8769  8784  8821  8836  8873  8888  8925  8940  8977  8992  9029  9044  9081  9096 609427 N/A N/A  8565  8580 GTGCAGGAGAGGCTGA  68 214  8617  8632  8669  8684  8721  8736  8773  8788  8825  8840  8877  8892  8929  8944  8981  8996  9033  9048  9085  9100 609428 N/A N/A  8567  8582 GAGTGCAGGAGAGGCT  69 215  8619  8634  8671  8686  8723  8738  8775  8790  8827  8842  8879  8894  8931  8946  8983  8998  9035  9050  9087  9102 609429 N/A N/A  8568  8583 GGAGTGCAGGAGAGGC  84 216  8620  8635  8672  8687  8724  8739  8776  8791  8828  8843  8880  8895  8932  8947  8984  8999  9036  9051  9088  9103 609430 N/A N/A  8572  8587 TAAAGGAGTGCAGGAG 107 217  8624  8639  8676  8691  8728  8743  8780  8795  8832  8847  8884  8899  8936  8951  8988  9003  9040  9055  9092  9107 609431 N/A N/A  8573  8588 GTAAAGGAGTGCAGGA  91 218  8625  8640  8677  8692  8729  8744  8833  8848  8885  8900  8937  8952  8989  9004  9041  9056 609432 N/A N/A  8575  8590 GGGTAAAGGAGTGCAG  81 219  8627  8642  8679  8694  8731  8746  8835  8850  8887  8902  8939  8954  8991  9006  9043  9058 609433 N/A N/A  8590  8605 CTGGCATCGAGACGGG  53 220  8642  8657  8694  8709  8798  8813  8954  8969  9006  9021 609434 N/A N/A  8593  8608 GCACTGGCATCGAGAC  43 221  8645  8660  8697  8712  8807  8816  8957  8972  9009  9024 609435 N/A N/A  8596  8611 GAAGCACTGGCATCGA  56 222  8648  8663  8700  8715  8804  8819  8960  8975  9012  9027 609436 N/A N/A 18852 18867 CCTGAATTTACAGCAT  60 223

Example 3: Effect of 4-8-4 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 20,000 cells per well were transfected using electroporation with 4,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Table 4 are 4-8-4 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises eight 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising four cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkkddddddddkkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Table 4 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 4 Percent control of human IRF4 mRNA with 4-8-4 cEt gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609518 156 171 5183 5198 GCTCATGCCGAACTCT 70 72 609519 213 228 5240 5255 GCTGTCGATCTGGTCG 94 73 609520 216 231 5243 5258 GCCGCTGTCGATCTGG 85 74 609521 219 234 5246 5261 CTTGCCGCTGTCGATC 58 75 609522 222 237 5249 5264 GTACTTGCCGCTGTCG 64 76 609523 238 253 5265 5280 CCCACACCAGCCCGGG 106 77 609524 241 256 5268 5283 TCTCCCACACCAGCCC 75 78 609525 244 259 5271 5286 CGTTCTCCCACACCAG 101 79 609526 294 309 5321 5336 GTCCTGCTTGCCCGCG 86 80 609527 297 312 5324 5339 GTAGTCCTGCTTGCCC 89 81 609528 334 349 N/A N/A CCCAAGCCTTGAAGAG 78 82 609529 431 446 6910 6925 AAGTCATTGCTCTTGT 72 83 609530 434 449 6913 6928 TCAAAGTCATTGCTCT 43 84 609531 437 452 6916 6931 TCCTCAAAGTCATTGC 90 85 609532 496 511 6975 6990 GAACAATCCTGTACAC 100 86 609533 514 529 6993 7008 CTTTTTTGGCTCCCTC 54 87 609534 517 532 N/A N/A CTCCTTTTTTGGCTCC 93 88 609535 610 625 N/A N/A GAACCTGCTGGGCTGG 87 89 609536 730 745 9220 9235 CTTGCCAGTGGTGGCC 109 90 609537 733 748 9223 9238 GGCCTTGCCAGTGGTG 94 91 609538 752 767 N/A N/A CAACCATTTTCACAAG 95 92 609539 755 770 N/A N/A TGGCAACCATTTTCAC 93 93 609540 758 773 N/A N/A ACCTGGCAACCATTTT 81 94 609541 761 776 N/A N/A GTCACCTGGCAACCAT 59 95 609542 764 779 10829 10844 CCTGTCACCTGGCAAC 63 96 609543 767 782 10832 10847 GTTCCTGTCACCTGGC 88 97 609544 770 785 10835 10850 AAGGTTCCTGTCACCT 86 98 609545 773 788 10838 10853 TAAAAGGTTCCTGTCA 57 99 609546 776 791 10841 10856 GCATAAAAGGTTCCTG 53 100 609547 780 795 10845 10860 ACAAGCATAAAAGGTT 60 101 609548 799 814 10864 10879 CCTGGGACTCAGGTGG 93 102 609549 802 817 10867 10882 GAGCCTGGGACTCAGG 81 103 609550 832 847 10897 10912 ACCTTATGCTTGGCTC 70 104 609551 835 850 10900 10915 CAGACCTTATGCTTGG 74 105 609554 943 958 13496 13511 GGGAGATCCGGCAGCC 113 106 609555 979 994 13532 13547 CCTGGTCCAGGTTGCT 66 107 609556 982 997 13535 13550 GGACCTGGTCCAGGTT 128 108 609557 985 1000 13538 13553 ACAGGACCTGGTCCAG 93 109 609558 1046 1061 13599 13614 TCCAGGTGGCTCAGCA 66 110 609559 1049 1064 13602 13617 CTCTCCAGGTGGCTCA 88 111 609560 1052 1067 13605 13620 CCCCTCTCCAGGTGGC 101 112 609561 1194 1209 13747 13762 TGTGTCAAAGAGCTTG 74 113 609562 1198 1213 13751 13766 GCTGTGTGTCAAAGAG 60 114 609563 1201 1216 13754 13769 ACTGCTGTGTGTCAAA 81 115 609564 1264 1279 17057 17072 GAGTCACCTGGAATCT 94 116 609565 1291 1306 17084 17099 GGTCTGGAAACTCCTC 69 117 609566 1294 1309 17087 17102 GAGGGTCTGGAAACTC 92 118 609567 1297 1312 17090 17105 TCTGAGGGTCTGGAAA 97 119 609568 1321 1336 17114 17129 GAGCTGTGATGAGCTT 95 120 609569 1342 1357 19459 19474 TGGCTAGCAGAGGTTC 91 121 609570 1345 1360 19462 19477 GTCTGGCTAGCAGAGG 91 122 609571 1348 1363 19465 19480 GTTGTCTGGCTAGCAG 48 123 609572 1389 1404 19506 19521 CCTCAGGAAATGTCCA 90 124 609574 2068 2083 20185 20200 CTTCCCTGAGAAATGG 64 126 609575 2071 2086 20188 20203 TTACTTCCCTGAGAAA 86 127 609576 2719 2734 20836 20851 AATAAGCAGAAACAGC 61 128 609577 3533 3548 21650 21665 ATCTATAAGATGTATA 64 129 609578 3536 3551 21653 21668 TGCATCTATAAGATGT 90 130 609579 3803 3818 21920 21935 TCCAGAAAATTCAGCT 74 131 609580 3809 3824 21926 21941 GCATTTTCCAGAAAAT 63 132 609581 N/A N/A 6995 7010 ACCTTTTTTGGCTCCC 81 133 609582 N/A N/A 7740 7755 ATTCTTAGAATGCATA 67 134 609583 N/A N/A 8504 8519 CAAACTCCTCAGGGAA 83 135 609584 N/A N/A 9242 9257 TTACCATTTTCACAAG 93 136 609585 N/A N/A 10796 10811 AATCAGATGATGTCTA 113 137 609586 N/A N/A 10813 10828 CTGCATTTGCAAATAA 108 138 609587 N/A N/A 10819 10834 GGCAACCTGCATTTGC 112 139 609588 N/A N/A 10825 10840 TCACCTGGCAACCTGC 72 140 609589 N/A N/A 11951 11966 CACCCACACAAGTCTT 86 141 609590 N/A N/A 11972 11987 GTTTATTTCCTACTCT 56 142 609591 N/A N/A 11978 11993 ATAGCTGTTTATTTCC 41 143 609592 N/A N/A 11985 12000 GATATAAATAGCTGTT 39 144 609593 N/A N/A 12024 12039 CATTTCATTTAATGTC 56 145 609594 N/A N/A 12031 12046 CAGGTTTCATTTCATT 46 146 609595 N/A N/A 13795 13810 CCTCCATTCTAACAGA 122 147

Example 4: Effect of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 20,000 cells per well were transfected using electroporation with 4,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 5 through 12 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 5 through 12 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 5 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 24 195 666225 5 20 3744 3759 TGAAACTGAGAGTGCG 81 224 666226 12 27 3751 3766 CGAGCGGTGAAACTGA 67 225 666227 19 34 3758 3773 CCAAGATCGAGCGGTG 63 226 666228 26 41 3765 3780 GTGGGTCCCAAGATCG 60 227 666229 40 55 3779 3794 AGCTGAGGGCAGCGGT 107 228 666230 47 62 3786 3801 GACTCGGAGCTGAGGG 77 229 666231 54 69 3793 3808 CGCCCTGGACTCGGAG 64 230 666232 61 76 N/A N/A CTGCACTCGCCCTGGA 58 231 666233 68 83 N/A N/A CTCTGCTCTGCACTCG 59 232 666234 75 90 5102 5117 CCGCCCGCTCTGCTCT 46 233 666235 82 97 5109 5124 GGGTCCTCCGCCCGCT 72 234 666236 101 116 5128 5143 CCGTCCGCGCCCGCGC 39 235 666237 129 144 5156 5171 GCCGCCCTCCAGGTTC 51 236 666238 136 151 5163 5178 CTCGGCCGCCGCCCTC 63 237 666239 143 158 5170 5185 TCTCCGCCTCGGCCGC 59 238 666240 150 165 5177 5192 GCCGAACTCTCCGCCT 49 239 666241 160 175 5187 5202 CCGCGCTCATGCCGAA 55 240 666242 167 182 5194 5209 CAGCTCACCGCGCTCA 58 241 666243 185 200 5212 5227 CGGAGCTTCCCGTTGC 65 242 666244 192 207 5219 5234 CCACTGGCGGAGCTTC 64 243 666245 304 319 5331 5346 CGCGGTTGTAGTCCTG 56 244 666246 311 326 5338 5353 TCCTCCTCGCGGTTGT 41 245 666247 327 342 5354 5369 CTTGAAGAGCGCGGCG 79 246 666248 338 353 N/A N/A AGTGCCCAAGCCTTGA 52 247 666249 348 363 6827 6842 TCCTTTAAACAGTGCC 50 248 666250 356 371 6835 6850 CGGAACTTTCCTTTAA 63 249 666251 363 378 6842 6857 GCCTTCTCGGAACTTT 68 250 666252 370 385 6849 6864 TGTCGATGCCTTCTCG 72 251 666253 377 392 6856 6871 TCCGGCTTGTCGATGC 59 252 666254 396 411 6875 6890 CGTCTTCCAGGTGGGA 84 253 666256 425 440 6904 6919 TTGCTCTTGTTCAAAG 71 254 666257 449 464 6928 6943 CGCTCAACCAGTTCCT 72 255 666258 456 471 6935 6950 CTGGCTCCGCTCAACC 48 256 666259 463 478 6942 6957 TGTCCAGCTGGCTCCG 61 257 666260 470 485 6949 6964 TCTGAGATGTCCAGCT 55 258 666261 484 499 6963 6978 ACACTTTGTACGGGTC 44 259 666262 507 522 6986 7001 GGCTCCCTCAGGAACA 81 260 666263 521 536 N/A N/A TTGGCTCCTTTTTTGG 87 261 666264 528 543 7846 7861 GAGCTGCTTGGCTCCT 81 262 666265 535 550 7853 7868 CCAGGGTGAGCTGCTT 69 263 666266 546 561 7864 7879 CTGCGGGTCCTCCAGG 80 264 666267 553 568 7871 7886 TGGACATCTGCGGGTC 56 265 666268 560 575 7878 7893 TGGCTCATGGACATCT 68 266 666269 577 592 7895 7910 TTGTCATGGTGTAGGG 59 267 666270 593 608 7911 7926 AGCGAAGGGTAAGGCG 86 268 666271 621 636 9111 9126 CATGTAGTTGTGAACC 73 269 666272 628 643 9118 9133 GTGGCATCATGTAGTT 73 270 666273 644 659 9134 9149 CAGCTTCGGTCGAGGG 42 271 666274 651 666 9141 9156 GTCCCTCCAGCTTCGG 97 272 666275 676 691 9166 9181 CCGGGTGTGGCTGATC 84 273 666276 695 710 9185 9200 GGACATTGGTACGGGA 50 274 666277 702 717 9192 9207 CGTCATGGGACATTGG 44 275 666278 725 740 9215 9230 CAGTGGTGGCCGCGGG 73 276 666279 740 755 9230 9245 CAAGCTGGGCCTTGCC 66 277 666280 747 762 9237 9252 ATTTTCACAAGCTGGG 61 278 666281 771 786 10836 10851 AAAGGTTCCTGTCACC 62 279 666282 775 790 10840 10855 CATAAAAGGTTCCTGT 91 280 666283 777 792 10842 10857 AGCATAAAAGGTTCCT 55 281 666284 779 794 10844 10859 CAAGCATAAAAGGTTC 71 282 666285 781 796 10846 10861 CACAAGCATAAAAGGT 74 283 666286 784 799 10849 10864 GGGCACAAGCATAAAA 80 284 666287 827 842 10892 10907 ATGCTTGGCTCTGTGG 24 285 666294 947 962 13500 13515 CCATGGGAGATCCGGC 46 286 666295 954 969 13507 13522 CGTATGTCCATGGGAG 12 287 666296 975 990 13528 13543 GTCCAGGTTGCTGGCG 47 288 666297 989 1004 13542 13557 GGGAACAGGACCTGGT 45 289 666298 1008 1023 13561 13576 GCCATTGTCCTCTGGG 75 290 666299 1015 1030 13568 13583 TCCTCTGGCCATTGTC 59 291 666300 1033 1048 13586 13601 GCAGCTTCTCAATGTT 57 292 666301 1042 1057 13595 13610 GGTGGCTCAGCAGCTT 63 293

TABLE 6 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 23 195 666302 1058 1073 13611 13626 ACCACGCCCCTCTCCA 91 294 666303 1065 1080 13618 13633 CCAGAGGACCACGCCC 72 295 666306 1101 1116 13654 13669 GCACAGTCTTTTCGCA 80 296 666307 1109 1124 13662 13677 CTGCTCTGGCACAGTC 17 297 666308 1116 1131 13669 13684 GTAGATCCTGCTCTGG 55 298 666309 1128 1143 13681 13696 GGGCCCGTCCCAGTAG 80 299 666311 1167 1182 13720 13735 TCTCTCCAGTTTGTTG 86 300 666312 1188 1203 13741 13756 AAAGAGCTTGCAGGTC 63 301 666313 1205 1220 13758 13773 AAGAACTGCTGTGTGT 88 302 666314 1212 1227 N/A N/A CTCTGACAAGAACTGC 70 303 666315 1219 1234 N/A N/A CTTGCAGCTCTGACAA 91 304 666317 1241 1256 17034 17049 GAGCGGCCGTGGTGAG 59 305 666318 1248 1263 17041 17056 TGGCAGGGAGCGGCCG 89 306 666319 1255 1270 17048 17063 GGAATCTTGGCAGGGA 52 307 666321 1275 1290 17068 17083 TCCAAAGCATAGAGTC 47 308 666322 1287 1302 17080 17095 TGGAAACTCCTCTCCA 84 309 666323 1311 1326 17104 17119 GAGCTTTCTTTGCCTC 87 310 666324 1338 1353 N/A N/A TAGCAGAGGTTCTACG 76 311 666326 1347 1362 19464 19479 TTGTCTGGCTAGCAGA 55 312 666327 1349 1364 19466 19481 AGTTGTCTGGCTAGCA 20 313 666328 1351 1366 19468 19483 ATAGTTGTCTGGCTAG 48 314 666329 1353 1368 19470 19485 ATATAGTTGTCTGGCT 59 315 666331 1381 1396 19498 19513 AATGTCCACTGTTTTG 53 316 666333 1417 1432 19534 19549 TGCTGATGTGTTCTGG 33 317 666335 1442 1457 19559 19574 ATAGATCTGTGGTAAT 62 318 666336 1449 1464 19566 19581 ATGGCGGATAGATCTG 43 319 666337 1456 1471 19573 19588 TAGAGGAATGGCGGAT 55 320 666338 1463 1478 19580 19595 TCTTGAATAGAGGAAT 58 321 666339 1485 1500 19602 19617 CACTCATCTTGACATT 64 322 666341 1538 1553 19655 19670 AAGACCCCGTATCCCC 58 323 666342 1686 1701 19803 19818 AAATCACTTGTCTTGG 51 324 666343 1715 1730 19832 19847 CTTTCACTAAAGTCAA 49 325 666344 1730 1745 19847 19862 GCAGTCAATTGGACGC 23 326 666345 1737 1752 19854 19869 TAAGAGGGCAGTCAAT 83 327 666346 1762 1777 19879 19894 TCCACTTCTGAATTCC 81 328 666347 1775 1790 19892 19907 CTGAACTGAAATCTCC 36 329 666348 1782 1797 19899 19914 TCAACCGCTGAACTGA 65 330 666349 1789 1804 19906 19921 ATTCTCCTCAACCGCT 65 331 666351 1803 1818 19920 19935 CTTGTCTCGCCGCAAT 35 332 666352 1810 1825 19927 19942 TTCCATGCTTGTCTCG 38 333 666353 1826 1841 19943 19958 TCAGATGTCACTGATT 65 334 666354 1840 1855 19957 19972 AGCTCATCTGCCAATC 67 335 666355 1848 1863 19965 19980 TTGAAATAAGCTCATC 67 336 666356 1885 1900 20002 20017 TCTACAGAACACAAGA 108 337 666357 1892 1907 20009 20024 ATGGCAGTCTACAGAA 102 338 666358 1899 1914 20016 20031 ATCAATGATGGCAGTC 43 339 666359 1908 1923 20025 20040 ACAGTGATCATCAATG 44 340 666360 1915 1930 20032 20047 AATTTTCACAGTGATC 66 341 666361 1922 1937 20039 20054 CTTGGTCAATTTTCAC 47 342 666362 1929 1944 20046 20061 CACATCACTTGGTCAA 28 343 666363 1956 1971 20073 20088 TAAAGAGCGCATTTCA 59 344 666364 1963 1978 20080 20095 AACAAATTAAAGAGCG 77 345 666365 1972 1987 20089 20104 CTAATCTACAACAAAT 77 346 666366 2000 2015 20117 20132 GCAAGTTTTCTCTGTC 44 347 666368 2023 2038 20140 20155 CTAGTCAGTGTCAATA 49 348 666369 2030 2045 20147 20162 CATCACTCTAGTCAGT 41 349 666370 2037 2052 20154 20169 AAGCAGTCATCACTCT 62 350 666371 2053 2068 20170 20185 GCACAGACATACCTAC 54 351 666373 2082 2097 20199 20214 CAATTTACATCTTACT 79 352 666374 2089 2104 20206 20221 GCTTCTTCAATTTACA 55 353 666375 2130 2145 20247 20262 GCAGCTCCTACATACA 47 354 666376 2138 2153 20255 20270 CAAGAACTGCAGCTCC 53 355 666377 2146 2161 20263 20278 GTCTTCCACAAGAACT 53 356 666378 2153 2168 20270 20285 AGCAAGTGTCTTCCAC 25 357

TABLE 7 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 23 195 666379 2161 2176 20278 20293 CTTCACTCAGCAAGTG 57 358 666380 2179 2194 20296 20311 CAGTCAAAGATTCATT 60 359 666381 2193 2208 20310 20325 TACAGGCACGGCTTCA 42 360 666382 2200 2215 20317 20332 CCAAGGCTACAGGCAC 37 361 666383 2207 2222 20324 20339 GGCCTCCCCAAGGCTA 82 362 666384 2235 2250 20352 20367 CCAGGAAACCGCTGGC 62 363 666385 2243 2258 20360 20375 GACCCACACCAGGAAA 98 364 666386 2250 2265 20367 20382 GCAGAGGGACCCACAC 79 365 666387 2308 2323 20425 20440 TTACTAACTGGCTTCC 51 366 666388 2315 2330 20432 20447 AGGAAGTTTACTAACT 42 367 666389 2328 2343 20445 20460 GACTCAAGAAAATAGG 36 368 666390 2335 2350 20452 20467 GTTTTTTGACTCAAGA 33 369 666392 2355 2370 20472 20487 CATCCAAGAGTAGCGC 27 370 666393 2376 2391 20493 20508 GTAGGACAGACAAAAA 77 371 666394 2383 2398 20500 20515 CTAGATTGTAGGACAG 52 372 666395 2390 2405 20507 20522 GACATTACTAGATTGT 30 373 666396 2397 2412 20514 20529 TTACTTAGACATTACT 52 374 666397 2404 2419 20521 20536 TTAACCATTACTTAGA 55 375 666398 2435 2450 20552 20567 GAGGGTCAAAAAGATG 67 376 666399 2450 2465 20567 20582 GCATCTCTAAAGAATG 39 377 666400 2461 2476 20578 20593 GAAGAATTTTAGCATC 44 378 666401 2468 2483 20585 20600 TTTATGCGAAGAATTT 67 379 666402 2475 2490 20592 20607 CTTCTTCTTTATGCGA 34 380 666403 2498 2513 20615 20630 TTAAGATTTATGTTCC 36 381 666404 2514 2529 20631 20646 GGCAACAGTTCAAGTA 43 382 666405 2521 2536 20638 20653 ACAGAAGGGCAACAGT 57 383 666406 2528 2543 20645 20660 TACTTGGACAGAAGGG 38 384 666407 2535 2550 20652 20667 AGTTAAGTACTTGGAC 58 385 666408 2542 2557 20659 20674 AACAGATAGTTAAGTA 81 386 666409 2578 2593 20695 20710 GCCAAACAAACAGAGG 78 387 666410 2585 2600 20702 20717 CTGGACAGCCAAACAA 79 388 666411 2592 2607 20709 20724 CTGATCGCTGGACAGC 80 389 666412 2599 2614 20716 20731 GCCATGGCTGATCGCT 70 390 666413 2606 2621 20723 20738 TAGTGTCGCCATGGCT 31 391 666414 2613 2628 20730 20745 CCTCCTTTAGTGTCGC 35 392 666415 2621 2636 20738 20753 CGGCTCCTCCTCCTTT 89 393 666416 2628 2643 20745 20760 GAGTCCCCGGCTCCTC 75 394 666417 2652 2667 20769 20784 TCCTGGCAGTGCTCTC 40 395 666418 2659 2674 20776 20791 TGGTGGGTCCTGGCAG 77 396 666420 2673 2688 20790 20805 CATCCTGCTTCCAGTG 70 397 666421 2681 2696 20798 20813 GTCAGCTCCATCCTGC 48 398 666422 2688 2703 20805 20820 TTCCGTAGTCAGCTCC 32 399 666423 2698 2713 20815 20830 GAGTGTGCAGTTCCGT 33 400 666424 2705 2720 20822 20837 GCCCACTGAGTGTGCA 90 401 666425 2714 2729 20831 20846 GCAGAAACAGCCCACT 59 402 666426 2737 2752 20854 20869 GAAGCATAGAACAGAT 49 403 666427 2744 2759 20861 20876 GCACGAGGAAGCATAG 42 404 666428 2749 2764 20866 20881 AATTGGCACGAGGAAG 72 405 666429 2751 2766 20868 20883 ATAATTGGCACGAGGA 33 406 666430 2753 2768 20870 20885 CTATAATTGGCACGAG 37 407 666431 2755 2770 20872 20887 AACTATAATTGGCACG 27 408 666432 2757 2772 20874 20889 CAAACTATAATTGGCA 47 409 666433 2759 2774 20876 20891 GTCAAACTATAATTGG 39 410 666434 2765 2780 20882 20897 GGCCCTGTCAAACTAT 60 411 666435 2772 2787 20889 20904 ATTTTAAGGCCCTGTC 66 412 666436 2779 2794 20896 20911 CCAAGTAATTTTAAGG 66 413 666437 2793 2808 20910 20925 GCATTTGGAAAAAGCC 29 414 666438 2810 2825 20927 20942 GGATTCTATAAATAGA 75 415 666439 2820 2835 20937 20952 GAGGTCTTTGGGATTC 26 416 666440 2827 2842 20944 20959 GCAAGTGGAGGTCTTT 26 417 666441 2834 2849 20951 20966 TACTTAAGCAAGTGGA 25 418 666442 2841 2856 20958 20973 ATAGGTATACTTAAGC 20 419 666443 2848 2863 20965 20980 GTAAGTGATAGGTATA 24 420 666444 2872 2887 20989 21004 GTACTTTCTCAAAACC 30 421 666445 2879 2894 20996 21011 TACTGCTGTACTTTCT 57 422 666446 2886 2901 21003 21018 CCCAGTCTACTGCTGT 34 423 666447 2902 2917 21019 21034 GGCCTGGAGGTGACGC 59 424 666448 2909 2924 21026 21041 GAGAAACGGCCTGGAG 56 425 666449 2916 2931 21033 21048 GTAGTATGAGAAACGG 17 426 666450 2923 2938 21040 21055 ATATCCTGTAGTATGA 43 427 666451 2930 2945 21047 21062 ATAGTAAATATCCTGT 64 428 666452 2940 2955 21057 21072 CCTGGGAGTAATAGTA 47 429 666453 2947 2962 21064 21079 TGCTGATCCTGGGAGT 21 430 666454 2954 2969 21071 21086 AATCTTCTGCTGATCC 28 431 666455 2961 2976 21078 21093 GCTACGCAATCTTCTG 42 432

TABLE 8 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 20 195 666457 2975 2990 21092 21107 ACACACATTTGAGAGC 34 433 666458 2992 3007 21109 21124 CCATTAGAAAAGCAGG 20 434 666459 3021 3036 21138 21153 TAGGTGCTTGTTGAAT 50 435 666460 3031 3046 21148 21163 AGGCACTTACTAGGTG 46 436 666461 3039 3054 21156 21171 GATACAGCAGGCACTT 37 437 666462 3046 3061 21163 21178 ATGTAGGGATACAGCA 26 438 666463 3053 3068 21170 21185 CTGTGTAATGTAGGGA 58 439 666464 3060 3075 21177 21192 GGCTGAACTGTGTAAT 28 440 666465 3067 3082 21184 21199 TGATAAAGGCTGAACT 59 441 666466 3074 3089 21191 21206 CTAAGCTTGATAAAGG 46 442 666467 3085 3100 21202 21217 CTCACTGCTCACTAAG 36 443 666468 3092 3107 21209 21224 TTCAGTGCTCACTGCT 31 444 666469 3099 3114 21216 21231 ATAATGTTTCAGTGCT 24 445 666470 3136 3151 21253 21268 CTGACTTTAATATTAG 44 446 666471 3178 3193 21295 21310 GTCTTTTCTGTAGTTA 10 447 666472 3192 3207 21309 21324 GTTCTCTACTGTTTGT 32 448 666473 3239 3254 21356 21371 GGAGAATTTGGGCTGG 25 449 666474 3246 3261 21363 21378 TTAGAGAGGAGAATTT 65 450 666475 3253 3268 21370 21385 GACACTTTTAGAGAGG 13 451 666476 3260 3275 21377 21392 TCTTGTGGACACTTTT 35 452 666477 3267 3282 21384 21399 CACCCCTTCTTGTGGA 60 453 666478 3274 3289 21391 21406 GAATAAACACCCCTTC 56 454 666479 3288 3303 21405 21420 GAAATGTGTTGGAAGA 32 455 666480 3335 3350 21452 21467 ACTCCATGAGGTTTTC 27 456 666481 3337 3352 21454 21469 TGACTCCATGAGGTTT 35 457 666482 3339 3354 21456 21471 GATGACTCCATGAGGT 33 458 666483 3341 3356 21458 21473 AAGATGACTCCATGAG 36 459 666485 3355 3370 21472 21487 ATGAAAGTGTGTGCAA 36 460 666486 3362 3377 21479 21494 GCACTGCATGAAAGTG 60 461 666487 3371 3386 21488 21503 CTACAAAGAGCACTGC 47 462 666488 3378 3393 21495 21510 CTGTTAGCTACAAAGA 44 463 666489 3385 3400 21502 21517 ATCTTCACTGTTAGCT 26 464 666490 3392 3407 21509 21524 GAGGTAAATCTTCACT 30 465 666491 3399 3414 21516 21531 GCAGAACGAGGTAAAT 38 466 666492 3406 3421 21523 21538 CCTCTGAGCAGAACGA 24 467 666493 3413 3428 21530 21545 AGCAAGGCCTCTGAGC 45 468 666494 3420 3435 21537 21552 GCTCCACAGCAAGGCC 42 469 666495 3427 3442 21544 21559 CAGTGGAGCTCCACAG 59 470 666496 3432 3447 21549 21564 CATGGCAGTGGAGCTC 13 471 666497 3434 3449 21551 21566 TACATGGCAGTGGAGC 29 472 666498 3436 3451 21553 21568 GGTACATGGCAGTGGA 13 473 666499 3438 3453 21555 21570 TGGGTACATGGCAGTG 26 474 666500 3440 3455 21557 21572 ACTGGGTACATGGCAG 41 475 666501 3442 3457 21559 21574 CTACTGGGTACATGGC 16 476 666502 3448 3463 21565 21580 CAAACCCTACTGGGTA 68 477 666503 3455 3470 21572 21587 GAAATGTCAAACCCTA 29 478 666504 3462 3477 21579 21594 GGCTAATGAAATGTCA 36 479 666505 3469 3484 21586 21601 GTTGCATGGCTAATGA 39 480 666506 3476 3491 21593 21608 TATCCATGTTGCATGG 39 481 666507 3491 3506 21608 21623 CTGCTGCCCAATACAT 53 482 666508 3498 3513 21615 21630 ACACAGTCTGCTGCCC 49 483 666509 3505 3520 21622 21637 TCACGAAACACAGTCT 32 484 666510 3512 3527 21629 21644 CTGCAGTTCACGAAAC 62 485 666511 3519 3534 21636 21651 TACATCACTGCAGTTC 32 486 666512 3526 3541 21643 21658 AGATGTATACATCACT 24 487 666513 3548 3563 21665 21680 CCCAAAATACTTTGCA 36 488 666514 3558 3573 21675 21690 GATAATATACCCCAAA 67 489 666515 3565 3580 21682 21697 CCCTTAGGATAATATA 51 490 666516 3572 3587 21689 21704 TTATCTTCCCTTAGGA 72 491 666517 3599 3614 21716 21731 GTGAAACAGCAGTTCT 38 492 666518 3606 3621 21723 21738 GGGCCCCGTGAAACAG 67 493 666519 3613 3628 21730 21745 CAGGTAAGGGCCCCGT 30 494 666520 3620 3635 21737 21752 AGGGTCACAGGTAAGG 32 495 666521 3627 3642 21744 21759 AGCAAAGAGGGTCACA 32 496 666522 3642 3657 21759 21774 GGTTAAATATTCTTCA 52 497 666523 3661 3676 21778 21793 TCTTTGAAGTGCTGTG 43 498 666524 3668 3683 21785 21800 GACAGCTTCTTTGAAG 64 499 666525 3675 3690 21792 21807 CTTCCAAGACAGCTTC 37 500 666526 3682 3697 21799 21814 AGACAGACTTCCAAGA 64 501 666527 3689 3704 21806 21821 GCTCCTGAGACAGACT 44 502 666528 3696 3711 21813 21828 ACAGGGTGCTCCTGAG 77 503 666529 3710 3725 21827 21842 GGAGAATTAAGAAGAC 68 504 666530 3721 3736 21838 21853 AGCATCCGCTTGGAGA 63 505 666531 3728 3743 21845 21860 GAAATGGAGCATCCGC 68 506 666532 3735 3750 21852 21867 AGCAATTGAAATGGAG 78 507

TABLE 9 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 20 195 666533 3742 3757 21859 21874 GTCACAAAGCAATTGA 33 508 666534 3786 3801 21903 21918 CACTGTTAAAGCAGCA 19 509 666535 3793 3808 21910 21925 TCAGCTCCACTGTTAA 27 510 666537 3824 3839 21941 21956 GGCCCCAGCCAAGAAG 92 511 666538 3831 3846 21948 21963 AGGTAGTGGCCCCAGC 40 512 666539 3865 3880 21982 21997 GTCAACATACACATAG 42 513 666540 3886 3901 22003 22018 GATCACTCAGAATTTT 50 514 666541 3893 3908 22010 22025 TACCCTGGATCACTCA 35 515 666542 3900 3915 22017 22032 TAGGTCATACCCTGGA 28 516 666543 3907 3922 22024 22039 CATTCCCTAGGTCATA 46 517 666544 3915 3930 22032 22047 AGCTAGTTCATTCCCT 52 518 666545 3922 3937 22039 22054 ATTTCATAGCTAGTTC 56 519 666546 3929 3944 22046 22061 CCTGAGTATTTCATAG 58 520 666547 3936 3951 22053 22068 TCCTAACCCTGAGTAT 73 521 666548 3950 3965 22067 22082 ACAAGTGCTAGGATTC 33 522 666549 3957 3972 22074 22089 TCCTGAGACAAGTGCT 45 523 666550 3964 3979 22081 22096 TTCAGAGTCCTGAGAC 54 524 666551 3968 3983 22085 22100 CCTTTTCAGAGTCCTG 42 525 666552 3972 3987 22089 22104 CGTTCCTTTTCAGAGT 70 526 666553 3974 3989 22091 22106 GCCGTTCCTTTTCAGA 38 527 666554 3976 3991 22093 22108 AAGCCGTTCCTTTTCA 71 528 666555 3982 3997 22099 22114 ATGAGGAAGCCGTTCC 42 529 666556 3996 4011 22113 22128 TATCAAGACAAGGAAT 93 530 666557 4003 4018 22120 22135 TCCACTTTATCAAGAC 35 531 666559 4017 4032 22134 22149 TCTAGTTTGCCAATTC 38 532 666560 4024 4039 22141 22156 ACTAAATTCTAGTTTG 100 533 666561 4035 4050 22152 22167 ACTGAGTACAAACTAA 68 534 666562 4042 4057 22159 22174 ACTGTCCACTGAGTAC 39 535 666563 4049 4064 22166 22181 CAACAGCACTGTCCAC 57 536 666564 4056 4071 22173 22188 AAATCTTCAACAGCAC 55 537 666565 4063 4078 22180 22195 AGTCCTCAAATCTTCA 46 538 666566 4071 4086 22188 22203 CTTTAACAAGTCCTCA 48 539 666567 4078 4093 22195 22210 CAGTGCTCTTTAACAA 63 540 666568 4085 4100 22202 22217 TATGACCCAGTGCTCT 60 541 666569 4093 4108 22210 22225 TTTTTCCATATGACCC 25 542 666570 4107 4122 22224 22239 GGAGACACATACATTT 78 543 666571 4117 4132 22234 22249 AATGCACCTGGGAGAC 38 544 666572 4124 4139 22241 22256 ACCAAGAAATGCACCT 43 545 666573 4134 4149 22251 22266 CAAGACATAAACCAAG 49 546 666574 4169 4184 22286 22301 CTTGAGGTTTTCCTAA 60 547 666575 4171 4186 22288 22303 TGCTTGAGGTTTTCCT 20 548 666576 4177 4192 22294 22309 AATTACTGCTTGAGGT 42 549 666577 4185 4200 22302 22317 GAGATATTAATTACTG 37 550 666578 4192 4207 22309 22324 GTTCCAGGAGATATTA 48 551 666579 4199 4214 22316 22331 CTATAGTGTTCCAGGA 34 552 666580 4206 4221 22323 22338 TGGTTCTCTATAGTGT 29 553 666581 4213 4228 22330 22345 GGTCACTTGGTTCTCT 21 554 666582 4220 4235 22337 22352 ATGAGTCGGTCACTTG 22 555 666583 4221 4236 22338 22353 AATGAGTCGGTCACTT 54 556 666584 4223 4238 22340 22355 TAAATGAGTCGGTCAC 24 557 666585 4225 4240 22342 22357 TGTAAATGAGTCGGTC 31 558 666586 4227 4242 22344 22359 GTTGTAAATGAGTCGG 13 559 666587 4229 4244 22346 22361 CAGTTGTAAATGAGTC 11 560 666588 4231 4246 22348 22363 TTCAGTTGTAAATGAG 44 561 666589 4237 4252 22354 22369 CTAGGTTTCAGTTGTA 46 562 666590 4244 4259 22361 22376 GGGCTTCCTAGGTTTC 61 563 666591 4272 4287 22389 22404 AACTCTCCTGTTTTCG 58 564 666592 4279 4294 22396 22411 GGCGACTAACTCTCCT 37 565 666593 4286 4301 22403 22418 TCTGTAGGGCGACTAA 35 566 666594 4294 4309 22411 22426 CTGGGTTTTCTGTAGG 52 567 666595 4301 4316 22418 22433 AGTCTAGCTGGGTTTT 37 568 666596 4308 4323 22425 22440 ACCCAATAGTCTAGCT 75 569 666598 4322 4337 22439 22454 TCTTTTTAGTTCATAC 74 570 666599 4330 4345 22447 22462 GCACAGTCTCTTTTTA 64 571 666600 4337 4352 22454 22469 CACCATGGCACAGTCT 21 572 666601 4344 4359 22461 22476 TTTTTCTCACCATGGC 26 573 666602 4365 4380 22482 22497 ATTTCACTGTAGGATT 62 574 666603 4372 4387 22489 22504 GCTGCTCATTTCACTG 59 575 666604 4379 4394 22496 22511 TGTAAGGGCTGCTCAT 51 576 666605 4386 4401 22503 22518 ACAATACTGTAAGGGC 25 577 666607 4407 4422 22524 22539 ACCTACCTGCCCTTGG 72 578 666608 4414 4429 22531 22546 CACTAATACCTACCTG 94 579 666609 4425 4440 22542 22557 GCTTTTTCAAACACTA 32 580

TABLE 10 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 25 195 666610 4434 4449 22551 22566 CAAAGACCAGCTTTTT 50 581 666611 4441 4456 22558 22573 CCTCGCTCAAAGACCA 50 582 666612 4448 4463 22565 22580 TTTATGCCCTCGCTCA 76 583 666613 4455 4470 22572 22587 AGCTGTATTTATGCCC 42 584 666614 4474 4489 22591 22606 TTGTTCCACCCCTGGG 53 585 666615 4483 4498 22600 22615 CTCCCAGAGTTGTTCC 66 586 666616 4491 4506 22608 22623 ACCCAAGACTCCCAGA 55 587 666617 4498 4513 22615 22630 TGCGAGTACCCAAGAC 41 588 666618 4505 4520 22622 22637 CAAGAGGTGCGAGTAC 78 589 666619 4520 4535 22637 22652 GAGCATCAACAAAGCC 42 590 666620 4527 4542 22644 22659 CCTGGCGGAGCATCAA 58 591 666621 4534 4549 22651 22666 TGGCCTTCCTGGCGGA 91 592 666622 4541 4556 22658 22673 ACACAAGTGGCCTTCC 69 593 666623 4575 4590 22692 22707 CTGAATTGTTACTAAA 79 594 666624 4582 4597 22699 22714 ACTGGATCTGAATTGT 43 595 666625 4589 4604 22706 22721 AGTTTACACTGGATCT 26 596 666626 4603 4618 22720 22735 GAGCAATGAACGGAAG 28 597 666627 4612 4627 22729 22744 GTGACTGGAGAGCAAT 26 598 666628 4641 4656 22758 22773 AACTTTCACCTGTGGG 50 599 666629 4669 4684 22786 22801 CCTTAACCAATCCCAA 57 600 666630 4676 4691 22793 22808 ATAAAGACCTTAACCA 73 601 666631 4686 4701 22803 22818 CGTAATACAAATAAAG 97 602 666632 4719 4734 22836 22851 GATGCAGCTGGCCACA 27 603 666633 4731 4746 22848 22863 ACCATTCAGACAGATG 70 604 666634 4738 4753 22855 22870 TTCACGCACCATTCAG 84 605 666635 4745 4760 22862 22877 GAGAGCCTTCACGCAC 77 606 666636 4752 4767 22869 22884 AAGGTCTGAGAGCCTT 83 607 666637 4759 4774 22876 22891 GGTGTGTAAGGTCTGA 59 608 666638 4766 4781 22883 22898 ACAAAATGGTGTGTAA 93 609 666639 4780 4795 22897 22912 GTAAAACATAACTTAC 92 610 666640 4807 4822 22924 22939 TCGAGATCAGTCTCAA 18 611 666641 4814 4829 22931 22946 ACCTGCATCGAGATCA 47 612 666642 4825 4840 22942 22957 CAAGGAGATCCACCTG 121 613 666643 4836 4851 22953 22968 TATCAGGATCTCAAGG 94 614 666644 4843 4858 22960 22975 AACAGGCTATCAGGAT 83 615 666645 4850 4865 22967 22982 TTCCTGTAACAGGCTA 23 616 666646 4866 4881 22983 22998 ACTGACCTTTACTTCA 57 617 666647 4898 4913 23015 23030 CCTCAAAGCTGTGAAA 80 618 666648 4905 4920 23022 23037 GCATGTTCCTCAAAGC 24 619 666649 4912 4927 23029 23044 TTCTTATGCATGTTCC 20 620 666650 4919 4934 23036 23051 GCTACATTTCTTATGC 80 621 666651 4927 4942 23044 23059 CTACTTCAGCTACATT 53 622 666652 4934 4949 23051 23066 GTCCCCTCTACTTCAG 59 623 666653 4949 4964 23066 23081 TGGCCCTTCTCTCACG 89 624 666654 4956 4971 23073 23088 GCCGGCCTGGCCCTTC 114 625 666655 4963 4978 23080 23095 TTGGCCTGCCGGCCTG 96 626 666656 4970 4985 23087 23102 AGGAGGGTTGGCCTGC 96 627 666657 4977 4992 23094 23109 CCATTGGAGGAGGGTT 68 628 666658 4982 4997 23099 23114 AATTTCCATTGGAGGA 67 629 666659 4984 4999 23101 23116 GGAATTTCCATTGGAG 56 630 666660 4986 5001 23103 23118 CGGGAATTTCCATTGG 50 631 666661 4988 5003 23105 23120 CACGGGAATTTCCATT 29 632 666662 4990 5005 23107 23122 AACACGGGAATTTCCA 27 633 666663 4992 5007 23109 23124 GCAACACGGGAATTTC 26 634 666664 5005 5020 23122 23137 GTCTCAGTTTGAAGCA 26 635 666665 5012 5027 23129 23144 CCCATCTGTCTCAGTT 34 636 666666 5019 5034 23136 23151 GTTAAGTCCCATCTGT 60 637 666667 5026 5041 23143 23158 ATTGCCTGTTAAGTCC 27 638 666668 5086 5101 23203 23218 GGCAGTTCTTTCAGCA 63 639 666669 5093 5108 23210 23225 ACCTGCTGGCAGTTCT 39 640 666670 5100 5115 23217 23232 GGGTCCTACCTGCTGG 64 641 666671 5124 5139 23241 23256 CAAGCTTTCATTTGGG 26 642 666672 5131 5146 23248 23263 GGAAATTCAAGCTTTC 36 643 666673 5147 5162 23264 23279 ACGCAGAGCCAGTAGG 50 644 666674 5149 5164 23266 23281 AAACGCAGAGCCAGTA 63 645 666675 5151 5166 23268 23283 CAAAACGCAGAGCCAG 42 646 666676 5171 5186 23288 23303 TCCTTTCCTACAGATC 67 647 666677 5179 5194 23296 23311 GTGAAGCATCCTTTCC 37 648 666678 5186 5201 23303 23318 TCAGTTTGTGAAGCAT 11 649 666679 5193 5208 23310 23325 ATCTACCTCAGTTTGT 58 650 666680 5200 5215 23317 23332 TAGCATTATCTACCTC 34 651 666681 5207 5222 23324 23339 GACAGCATAGCATTAT 23 652 666682 5214 5229 23331 23346 TACCAACGACAGCATA 39 653 666683 5221 5236 23338 23353 TGATGTATACCAACGA 13 654 666684 5246 5261 23363 23378 GCAGAGCAATTTACAT 34 655 666685 5253 5268 23370 23385 TTGCTTTGCAGAGCAA 72 656

TABLE 11 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 15 195 666688 N/A N/A 18773 18788 CTTAAGCTAAACTCCT 73 657 666689 N/A N/A 18780 18795 ATGACGACTTAAGCTA 38 658 666690 N/A N/A 18803 18818 ATATTAGAGACTGTAT 75 659 666691 N/A N/A 18817 18832 CTCCATCAATCATGAT 67 660 666692 N/A N/A 18824 18839 GAATGCTCTCCATCAA 55 661 666693 N/A N/A 18831 18846 GCAAGCTGAATGCTCT 47 662 666694 N/A N/A 18838 18853 ATCTAAGGCAAGCTGA 63 663 666695 N/A N/A 18845 18860 TTACAGCATCTAAGGC 79 664 666696 N/A N/A 18866 18881 AATGTTCAGCTTTTCC 37 665 666697 N/A N/A 18873 18888 ACGCCTTAATGTTCAG 23 666 666699 N/A N/A 22817 22832 GACTTCGGGAGATACG 86 667 666700 N/A N/A 22824 22839 CACAGAGGACTTCGGG 51 668 666701 N/A N/A 3917 3932 CCCTTCCCGCTCCGAA 84 669 666702 N/A N/A 4018 4033 AGGCTCGGGCAGAGCC 99 670 666703 N/A N/A 4254 4269 GAGCACCGCGCCGGAG 93 671 666704 N/A N/A 4288 4303 ATGTGGAGCTCCTCCT 80 672 666705 N/A N/A 4546 4561 GCCCCGAATAGGACCC 79 673 666706 N/A N/A 4593 4608 CCGGGTTCCTCCACCC 91 674 666707 N/A N/A 4653 4668 CCTGGCGGCCTCCACG 131 675 666709 N/A N/A 4856 4871 AGGCTCCTACCCGCCT 94 676 666710 N/A N/A 4965 4980 TGCCGTGTCAGGGTCG 89 677 666711 N/A N/A 4993 5008 GAGTCTTTGAGGCTGC 66 678 666712 N/A N/A 5759 5774 ACAGGCGCGGACGCAC 80 679 666713 N/A N/A 5867 5882 CGAGAAGAGAGGCTGA 93 680 666714 N/A N/A 6304 6319 GACACATTTTCTGGGT 36 681 666715 N/A N/A 7071 7086 GATGCTCAGAATGAAG 57 682 666716 N/A N/A 7190 7205 CTATGCTGAACCCCAC 99 683 666717 N/A N/A 7197 7212 AATTCTCCTATGCTGA 87 684 666718 N/A N/A 7380 7395 AGTGATGAGATATTCC 41 685 666719 N/A N/A 7519 7534 TCCAGATGCAGATTCC 54 686 666720 N/A N/A 7536 7551 TCACCTGAAGGATTTG 75 687 666721 N/A N/A 7731 7746 ATGCATAACACGGTGT 55 688 666722 N/A N/A 7963 7978 ACACAGCCTCGCCCTC 80 689 666723 N/A N/A 8094 8109 AGCACCGTGTGGAAAG 43 690 666724 N/A N/A 8126 8141 ACCCTCCCCAACTTAA 90 691 666725 N/A N/A 8336 8351 CTGGCACCAAAAGTAC 82 692 666726 N/A N/A 8539 8554 ACTGGCATTGAGACGG 21 693 8747 8762 8851 8866 8903 8918 9059 9074 666727 N/A N/A 8540 8555 CACTGGCATTGAGACG 24 694 8748 8763 8852 8867 8904 8919 9060 9075 666728 N/A N/A 8541 8556 GCACTGGCATTGAGAC 27 695 8749 8764 8853 8868 8905 8920 9061 9076 666729 N/A N/A 8543 8558 AAGCACTGGCATTGAG 58 696 8751 8766 8855 8870 8907 8922 9063 9078 666730 N/A N/A 8544 8559 GAAGCACTGGCATTGA 45 697 8752 8767 8856 8871 8908 8923 9064 9079 666731 N/A N/A 8545 8560 AGAAGCACTGGCATTG 55 698 9065 9080 666732 N/A N/A 8548 8563 ATAAGAAGCACTGGCA 49 699 8704 8719 8808 8823 8964 8979 9068 9083 666733 N/A N/A 8549 8564 GATAAGAAGCACTGGC 37 700 8705 8720 8809 8824 8965 8980 9069 9084 666734 N/A N/A 8551 8566 GAGATAAGAAGCACTG 56 701 8707 8722 8811 8826 8967 8982 9071 9086 666735 N/A N/A 8553 8568 CTGAGATAAGAAGCAC 63 702 8709 8724 8813 8828 8969 8984 9073 9088 666737 N/A N/A 8574 8589 GGTAAAGGAGTGCAGG 54 703 8626 8641 8678 8693 8730 8745 8834 8849 8886 8901 8938 8953 8990 9005 9042 9057 666738 N/A N/A 8591 8606 ACTGGCATCGAGACGG 21 704 8643 8658 8695 8710 8799 8814 8955 8970 9007 9022 666739 N/A N/A 8592 8607 CACTGGCATCGAGACG 39 705 8644 8659 8696 8711 8800 8815 8956 8971 9008 9023 666740 N/A N/A 8594 8609 AGCACTGGCATCGAGA 41 706 8646 8661 8698 8713 8802 8817 8958 8973 9010 9025 666741 N/A N/A 8595 8610 AAGCACTGGCATCGAG 73 707 8647 8662 8699 8714 8803 8818 8959 8974 9011 9026 666742 N/A N/A 8597 8612 GGAAGCACTGGCATCG 45 708 8649 8664 9013 9028 666743 N/A N/A 8600 8615 ATAGGAAGCACTGGCA 58 709 8652 8667 8756 8771 8860 8875 8912 8927 9016 9031 666744 N/A N/A 8601 8616 GATAGGAAGCACTGGC 38 710 8653 8668 8757 8772 8861 8876 8913 8928 9017 9032 666745 N/A N/A 8602 8617 AGATAGGAAGCACTGG 54 711 8654 8669 8758 8773 8862 8877 8914 8929 9018 9033 666746 N/A N/A 8603 8618 GAGATAGGAAGCACTG 52 712 8655 8670 8759 8774 8863 8878 8915 8930 9019 9034 666747 N/A N/A 8604 8619 TGAGATAGGAAGCACT 62 713 8656 8671 8760 8775 8864 8879 8916 8931 9020 9035 666748 N/A N/A 8605 8620 CTGAGATAGGAAGCAC 74 714 8657 8672 8761 8776 8865 8880 8917 8932 9021 9036 666749 N/A N/A 9410 9425 GGATGCCACCATCCCA 105 715 666750 N/A N/A 9490 9505 GGAGTGATCTCTGTGG 32 716 666751 N/A N/A 9708 9723 GTAGGTAGGCACCTGT 25 717 666752 N/A N/A 9796 9811 GAGGGAGCTCATTTTG 76 718 666753 N/A N/A 9960 9975 AAAGGCCAAATTGCAA 48 719 666754 N/A N/A 9997 10012 CTGCAGCCAAGGATAA 52 720 666818 N/A N/A 3925 3940 GGCGCGCTCCCTTCCC 90 721 666819 N/A N/A 4685 4700 CCCTTCCCCGCGACTC 81 722 666820 N/A N/A 4717 4732 TTGCCTTCGCTCACTC 78 723 666821 N/A N/A 5757 5772 AGGCGCGGACGCACGG 69 724 666822 N/A N/A 6997 7012 CTACCTTTTTTGGCTC 83 725 666823 N/A N/A 7585 7600 TGCCTTGTGACATAAA 55 726 666824 N/A N/A 8151 8166 ATATTCCAACAGGCGG 47 727 666825 N/A N/A 8437 8452 TGAACCCTTCATCAGA 52 728 666826 N/A N/A 9313 9328 AGACCAGGATTCGCCA 67 729

TABLE 12 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleotide linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  23 195 666755 N/A N/A 10003 10018 CACTACCTGCAGCCAA  69 730 666756 N/A N/A 10020 10035 AGAGTGGTACACCTCT  78 731 666757 N/A N/A 10256 10271 GTCTCTACCTACTCCA  59 732 666758 N/A N/A 10308 10323 AACACCTGATCTTGCT 110 733 666759 N/A N/A 10366 10381 ACCATGTTCCTGAAGA  56 734 666760 N/A N/A 10660 10675 CTCTTGTGAGGCAAAG  52 735 666761 N/A N/A 10712 10727 GCGCCCAATCACCTTC  97 736 666762 N/A N/A 11002 11017 ATCGAATCTGCCCAAA  74 737 666763 N/A N/A 11010 11025 AAAGTCCCATCGAATC  87 738 666764 N/A N/A 11024 11039 CAAAGCAAGTGTCTAA  77 739 666765 N/A N/A 11070 11085 CTCACACACAGGATGT  82 740 666767 N/A N/A 11344 11359 GAGGAGGAGGACTTAT  49 741 666768 N/A N/A 11408 11423 TATTACTCTTAGGCAC  56 742 666769 N/A N/A 11545 11560 GTGCCTTCTTTATAGT  51 743 666770 N/A N/A 11551 11566 CTAGAGGTGCCTTCTT  74 744 666771 N/A N/A 11682 11697 TGCCAGAGGGCAAGAT  98 745 666772 N/A N/A 11975 11990 GCTGTTTATTTCCTAC  34 746 666773 N/A N/A 11988 12003 CGAGATATAAATAGCT  42 747 666774 N/A N/A 11990 12005 GCCGAGATATAAATAG  60 748 666775 N/A N/A 12032 12047 GCAGGTTTCATTTCAT  56 749 666776 N/A N/A 12034 12049 GGGCAGGTTTCATTTC  68 750 666777 N/A N/A 12067 12082 AGAGTGGGTGTTGGCC  75 751 666779 N/A N/A 12274 12289 ATGGAACCCCAAAATC  76 752 666780 N/A N/A 12292 12307 GCTGCCACTGGTAACT  74 753 666781 N/A N/A 12374 12389 TCGCCCATGAGTTGAA  61 754 666782 N/A N/A 12516 12531 AGTTCATGTAAAGTCT  39 755 666783 N/A N/A 12865 12880 TCGGTCCACATACCTG  61 756 666784 N/A N/A 13199 13214 CTTGGAGAAGTCCCGT  61 757 666785 N/A N/A 13330 13345 CCTGTGGTGGAACTCT  73 758 666786 N/A N/A 14020 14035 AAGAGGCGACTGCTGA  65 759 666787 N/A N/A 14028 14043 ACTGTTTTAAGAGGCG  30 760 666788 N/A N/A 14046 14061 ATAGTGCAATGAAATG  64 761 666789 N/A N/A 14094 14109 GGGAACCTGAAAAGAG  79 762 666790 N/A N/A 14338 14353 GAGGTTCGCTGAATTG  70 763 666791 N/A N/A 14687 14702 ATCTGGATGAGCTTAC  62 764 666792 N/A N/A 14732 14747 AGCTTAGTTATCTGGG  41 765 666793 N/A N/A 15213 15228 GAACACCATGCCCAAC  93 766 666794 N/A N/A 15304 15319 GAGCTCTTCCAGATCC  95 767 666795 N/A N/A 15596 15611 TAGTCGCGCAAGTCTA 108 768 666796 N/A N/A 15777 15792 ATCCTTAATTATGCAG  61 769 666797 N/A N/A 15790 15805 GTGTTTGGTGGGAATC  54 770 666798 N/A N/A 15985 16000 CTAACTTACAGGACTA  69 771 666799 N/A N/A 16059 16074 TAGTGCTGTGCAGACC  68 772 666800 N/A N/A 16069 16084 GGGCTGTCCCTAGTGC 109 773 666801 N/A N/A 16171 16186 AATGTCACGCCCGCAA  81 774 666802 N/A N/A 16340 16355 GGCCTCCCGCTTGTGG 118 775 666803 N/A N/A 16383 16398 GAACAGTAACTTGACT  96 776 666804 N/A N/A 16419 16434 AACTCTGAGTAGACTT  72 777 666805 N/A N/A 16471 16486 GAGCACCAGCCATCGG  77 778 666806 N/A N/A 16649 16664 GACCAATTTATGCCAT  62 779 666807 N/A N/A 16664 16679 CACGCAATGGCAAAAG  81 780 666808 N/A N/A 16824 16839 TCCTTTGGGTGCTTTC  68 781 666809 N/A N/A 16888 16903 CTCTTACTCCGCTGAG 105 782 666810 N/A N/A 16953 16968 ACACACCAGGTTAAAC  88 783 666811 N/A N/A 17135 17150 AGTGTAACTGAGGACT 107 784 666812 N/A N/A 17740 17755 TGTTACTTGCCACAGT  61 785 666813 N/A N/A 17916 17931 ATGCAAGCCCGTTAAT  96 786 666814 N/A N/A 17966 17981 ATGAGAACTTTGATGT  85 787 666815 N/A N/A 18136 18151 AAGTCAATCACTGGAG  39 788 666816 N/A N/A 18196 18211 TTAGCAATTCCTGTTG  80 789 666817 N/A N/A 18938 18953 GCTAACTGGCCTCAAA  91 790 666827 N/A N/A 10288 10303 CTTCATTTACTGTTAC  69 791 666828 N/A N/A 10484 10499 CAACAGAGCTGAGAGT  80 792 666829 N/A N/A 11844 11859 CTCACGAGCACCTCAG  58 793 666830 N/A N/A 13244 13259 CTTGTCTCCCCCAGAG  97 794 666831 N/A N/A 13798 13813 CCACCTCCATTCTAAC 115 795 666832 N/A N/A 14081 14096 GAGCCGCCACATCAGC  77 796 666833 N/A N/A 14193 14208 CCACCCTCCGTCTCAC  67 797 666834 N/A N/A 14242 14257 TATAGCACTCTCCTAT  98 798 666836 N/A N/A 16203 16218 CAAGACAAATCTTCTG  87 799 666837 N/A N/A 16641 16656 TATGCCATGGACAAGT  81 800 666838 N/A N/A 16948 16963 CCAGGTTAAACAGGAA  94 801 666839 N/A N/A 19232 19247 GACTTAATTCTGGGTT  65 802 666840 N/A N/A 19412 19427 ACTGAGATATCCTGCA  81 803

Example 5: Effect of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 20,000 cells per well were transfected using electroporation with 4,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 13 through 24 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 13 through 24 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 13 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  24 195 881075    2   17  3741  3756 AACTGAGAGTGCGAGG  99 804 881099  342  357  6821  6836 AAACAGTGCCCAAGCC  75 805 881122  624  639  9114  9129 CATCATGTAGTTGTGA  59 806 881169 1192 1207 13745 13760 TGTCAAAGAGCTTGCA  89 807 881193 1416 1431 19533 19548 GCTGATGTGTTCTGGT  21 808 881217 1740 1755 19857 19872 CAGTAAGAGGGCAGTC  50 809 881241 1927 1942 20044 20059 CATCACTTGGTCAATT  71 810 881265 2042 2057 20159 20174 CCTACAAGCAGTCATC  46 811 881289 2194 2209 20311 20326 CTACAGGCACGGCTTC  48 812 881313 2385 2400 20502 20517 TACTAGATTGTAGGAC  73 813 881337 2512 2527 20629 20644 CAACAGTTCAAGTATT  65 814 881361 2665 2680 20782 20797 TTCCAGTGGTGGGTCC  71 815 881385 2826 2841 20943 20958 CAAGTGGAGGTCTTTG  33 816 881409 2911 2926 21028 21043 ATGAGAAACGGCCTGG  36 817 881433 3047 3062 21164 21179 AATGTAGGGATACAGC  38 818 881457 3273 3288 21390 21405 AATAAACACCCCTTCT  79 819 881481 3419 3434 21536 21551 CTCCACAGCAAGGCCT  49 820 881505 3507 3522 21624 21639 GTTCACGAAACACAGT  37 821 881528 3625 3640 21742 21757 CAAAGAGGGTCACAGG  37 822 881552 3903 3918 22020 22035 CCCTAGGTCATACCCT  43 823 881576 4052 4067 22169 22184 CTTCAACAGCACTGTC  44 824 881595 4285 4300 22402 22417 CTGTAGGGCGACTAAC  51 825 881619 4412 4427 22529 22544 CTAATACCTACCTGCC  53 826 881643 4545 4560 22662 22677 GCACACACAAGTGGCC  36 827 881667 4602 4617 22719 22734 AGCAATGAACGGAAGT  20 828 881691 4839 4854 22956 22971 GGCTATCAGGATCTCA  55 829 881715 5030 5045 23147 23162 CCCCATTGCCTGTTAA  80 830 881739 5206 5221 23323 23338 ACAGCATAGCATTATC  36 831 881763 N/A N/A 18778 18793 GACGACTTAAGCTAAA  38 832 881787 N/A N/A 18847 18862 ATTTACAGCATCTAAG  53 833 881809 N/A N/A  4075  4090 GCTCATCCCGTCCAGC  85 834 881832 N/A N/A  4402  4417 GGAGAGCGGAGGCGGG  79 835 881855 N/A N/A  4644  4659 CTCCACGCGCGGAGGA 101 836 881879 N/A N/A  5038  5053 GGGCACCCCGCCCCGA  99 837 881903 N/A N/A  5660  5675 GCACGGACGAACGCGC  77 838 881925 N/A N/A  5777  5792 ACGAAAACAGCCGCCG  86 839 881947 N/A N/A  6136  6151 GAGCAAATTGAGACCA  45 840 881971 N/A N/A  6415  6430 GCGCATAGGTCCTTCA  37 841 881993 N/A N/A  7155  7170 CCACATAACTCAGGCA  30 842 882017 N/A N/A  7376  7391 ATGAGATATTCCTCTC  68 843 882041 N/A N/A  7696  7711 CAGCAACTCCCTTGGG  77 844 882065 N/A N/A  8145  8160 CAACAGGCGGACACGC  58 845 882088 N/A N/A  8385  8400 ATGGAGATACTTGTAC  38 846 882112 N/A N/A  8554  8569 GCTGAGATAAGAAGCA  68 847  8710  8725  8814  8829  8970  8985  9074  9089 882135 N/A N/A  9362  9377 GCACACGCAGCCTCTA  95 848 882158 N/A N/A  9606  9621 CCACTATTCGAGAGAA  43 849 882182 N/A N/A  9981  9996 CCTGAACATGACTGGG  62 850 882205 N/A N/A 10162 10177 GAATTTCAGGAGCTAG  39 851 882229 N/A N/A 10314 10329 GCACAGAACACCTGAT  44 852 882253 N/A N/A 10616 10631 AGCTGATGGAGAAACG  90 853 882277 N/A N/A 11088 11103 CTAAATCACCCTGGTC  53 854 882301 N/A N/A 11366 11381 GAACTAATGTCCCCAG  39 855 882325 N/A N/A 11666 11681 GAGAATTCCAAACCTT  24 856 882349 N/A N/A 11940 11955 GTCTTAGGTGTTCAAG  44 857 882373 N/A N/A 12157 12172 CAGCAGGTTTTGAGAC  80 858 882397 N/A N/A 12512 12527 CATGTAAAGTCTGCTG  42 859 882420 N/A N/A 12893 12908 ATATAACGGTGTTTCA  42 860 882443 N/A N/A 13175 13190 ACCCATTTAATCTGTC  43 861 882465 N/A N/A 13791 13806 CATTCTAACAGATAAC 102 862 882487 N/A N/A 14150 14165 ACACACCTGACAACCA  64 863 882510 N/A N/A 14374 14389 GCATGACAGGGCGAGG  57 864 882533 N/A N/A 14708 14723 TGCTTTGGGCACCAAA  48 865 882557 N/A N/A 15460 15475 GCCCAGCAAGAGGCAC  93 866 882580 N/A N/A 15804 15819 TAGATAACATGAGAGT  50 867 882603 N/A N/A 15925 15940 CAAATGACTTAGTCAG  45 868 882627 N/A N/A 16168 16183 GTCACGCCCGCAAAAG  61 869 882650 N/A N/A 16388 16403 GGATGGAACAGTAACT  48 870 882674 N/A N/A 16667 16682 AGTCACGCAATGGCAA  50 871 882698 N/A N/A 16900 16915 GAGGAATGAGCACTCT  74 872 882722 N/A N/A 17238 17253 GGTTACGCTTATTTTT  36 873 882746 N/A N/A 17493 17508 GCTAGATAGCATTCTT  44 874 882770 N/A N/A 17733 17748 TGCCACAGTTGAACCC  50 875 882794 N/A N/A 17987 18002 TAGCATCAGAGCTAGA  51 876 882817 N/A N/A 18582 18597 CTGGATTGATGTGATA  57 877 882841 N/A N/A 18961 18976 ACTACTATTGTGGAAA  82 878 882864 N/A N/A 19234 19249 GTGACTTAATTCTGGG  49 879

TABLE 14 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  14 195 881076    6   21  3745  3760 GTGAAACTGAGAGTGC 120 880 881100  343  358  6822  6837 TAAACAGTGCCCAAGC  59 881 881123  682  697  9172  9187 GGATTTCCGGGTGTGG  39 882 881170 1206 1221 13759 13774 CAAGAACTGCTGTGTG 105 883 881194 1432 1447 19549 19564 GGTAATCTTCTGGATT  70 884 881218 1741 1756 19858 19873 ACAGTAAGAGGGCAGT  16 885 881242 1930 1945 20047 20062 ACACATCACTTGGTCA  22 886 881266 2043 2058 20160 20175 ACCTACAAGCAGTCAT  54 887 881290 2195 2210 20312 20327 GCTACAGGCACGGCTT  16 888 881314 2387 2402 20504 20519 ATTACTAGATTGTAGG  50 889 881338 2515 2530 20632 20647 GGGCAACAGTTCAAGT  64 890 881362 2686 2701 20803 20818 CCGTAGTCAGCTCCAT  27 891 881386 2828 2843 20945 20960 AGCAAGTGGAGGTCTT  37 892 881410 2912 2927 21029 21044 TATGAGAAACGGCCTG  55 893 881434 3048 3063 21165 21180 TAATGTAGGGATACAG  22 894 881458 3275 3290 21392 21407 AGAATAAACACCCCTT  26 895 881482 3429 3444 21546 21561 GGCAGTGGAGCTCCAC  27 896 881506 3510 3525 21627 21642 GCAGTTCACGAAACAC  20 897 881529 3628 3643 21745 21760 CAGCAAAGAGGGTCAC  39 898 881553 3904 3919 22021 22036 TCCCTAGGTCATACCC  42 899 881577 4058 4073 22175 22190 TCAAATCTTCAACAGC  24 900 881596 4300 4315 22417 22432 GTCTAGCTGGGTTTTC  37 901 881620 4413 4428 22530 22545 ACTAATACCTACCTGC  52 902 881644 4547 4562 22664 22679 ACGCACACACAAGTGG  41 903 881668 4608 4623 22725 22740 CTGGAGAGCAATGAAC  33 904 881692 4844 4859 22961 22976 TAACAGGCTATCAGGA  74 905 881716 5040 5055 23157 23172 GGGAAGTGGACCCCAT  91 906 881740 5208 5223 23325 23340 CGACAGCATAGCATTA  25 907 881764 N/A N/A 18782 18797 ATATGACGACTTAAGC  62 908 881788 N/A N/A 18849 18864 GAATTTACAGCATCTA  24 909 881810 N/A N/A  4084  4099 GTCCGGTTAGCTCATC  55 910 881833 N/A N/A  4403  4418 GGGAGAGCGGAGGCGG  78 911 881856 N/A N/A  4703  4718 TCCCAACCCGCTTCTC  86 912 881880 N/A N/A  5056  5071 CACGAGGCACCGCACT 113 913 881904 N/A N/A  5664  5679 GAACGCACGGACGAAC  82 914 881926 N/A N/A  5778  5793 GACGAAAACAGCCGCC  38 915 881948 N/A N/A  6141  6156 TCTGAGAGCAAATTGA  71 916 881994 N/A N/A  7157  7172 ACCCACATAACTCAGG  69 917 882018 N/A N/A  7384  7399 GCATAGTGATGAGATA  41 918 882042 N/A N/A  7717  7732 GTTGACAAATAAGGTA  40 919 882066 N/A N/A  8146  8161 CCAACAGGCGGACACG  17 920 882089 N/A N/A  8391  8406 AGGACAATGGAGATAC  28 921 882113 N/A N/A  8566  8581 AGTGCAGGAGAGGCTG  64 922  8618  8633  8670  8685  8722  8737  8774  8789  8826  8841  8878  8893  8930  8945  8982  8997  9034  9049  9086  9101 882136 N/A N/A  9386  9401 AGAAAACCCCCCTCTC  77 923 882159 N/A N/A  9608  9623 CACCACTATTCGAGAG  52 924 882183 N/A N/A  9986 10001 GATAACCTGAACATGA  83 925 882206 N/A N/A 10172 10187 ACGCAAGTCTGAATTT  35 926 882230 N/A N/A 10352 10367 GACAATGTTGTATGAA  28 927 882254 N/A N/A 10634 10649 GACCAACTGGAAAACC  44 928 882278 N/A N/A 11089 11104 CCTAAATCACCCTGGT  73 929 882302 N/A N/A 11368 11383 CAGAACTAATGTCCCC  31 930 882326 N/A N/A 11669 11684 GATGAGAATTCCAAAC  16 931 882350 N/A N/A 11950 11965 ACCCACACAAGTCTTA 104 932 882374 N/A N/A 12161 12176 TCAACAGCAGGTTTTG  59 933 882398 N/A N/A 12515 12530 GTTCATGTAAAGTCTG  14 934 882421 N/A N/A 12896 12911 AAAATATAACGGTGTT  69 935 882444 N/A N/A 13196 13211 GGAGAAGTCCCGTGGA  41 936 882466 N/A N/A 13881 13896 CGCCGAAGTCAACAGG  56 937 882488 N/A N/A 14160 14175 CATGAGGGTGACACAC  53 938 882511 N/A N/A 14376 14391 CAGCATGACAGGGCGA  44 939 882534 N/A N/A 14716 14731 GACATATTTGCTTTGG  42 940 882558 N/A N/A 15497 15512 AGCTTAGTCACCACGG  33 941 882581 N/A N/A 15805 15820 CTAGATAACATGAGAG  47 942 882604 N/A N/A 15926 15941 CCAAATGACTTAGTCA  49 943 882628 N/A N/A 16177 16192 CAATAAAATGTCACGC  53 944 882651 N/A N/A 16390 16405 AGGGATGGAACAGTAA  53 945 882675 N/A N/A 16691 16706 CGGGAGATAAAGAACA  69 946 882699 N/A N/A 16905 16920 TGCAAGAGGAATGAGC  16 947 882723 N/A N/A 17239 17254 GGGTTACGCTTATTTT  35 948 882747 N/A N/A 17508 17523 AATGAAGATCCACTAG  39 949 882771 N/A N/A 17746 17761 TTTTAGTGTTACTTGC  68 950 882795 N/A N/A 17990 18005 AAATAGCATCAGAGCT  87 951 882818 N/A N/A 18584 18599 AACTGGATTGATGTGA  23 952 882842 N/A N/A 18964 18979 AAAACTACTATTGTGG  86 953 882865 N/A N/A 19240 19255 GCCAAAGTGACTTAAT  30 954 882898 N/A N/A  6422  6437 AAGAATGGCGCATAGG  19 955

TABLE 15 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  45  195 881077    7   22  3746  3761 GGTGAAACTGAGAGTG  92  956 881101  344  359  6823  6838 TTAAACAGTGCCCAAG  79  957 881124  689  704  9179  9194 TGGTACGGGATTTCCG  63  958 881171 1207 1222 13760 13775 ACAAGAACTGCTGTGT  93  959 881195 1433 1448 19550 19565 TGGTAATCTTCTGGAT  76  960 881219 1744 1759 19861 19876 AAAACAGTAAGAGGGC  61  961 881243 1934 1949 20051 20066 GTAAACACATCACTTG  49  962 881267 2044 2059 20161 20176 TACCTACAAGCAGTCA  44  963 881291 2196 2211 20313 20328 GGCTACAGGCACGGCT  58  964 881315 2388 2403 20505 20520 CATTACTAGATTGTAG  91  965 881339 2520 2535 20637 20652 CAGAAGGGCAACAGTT  62  966 881363 2701 2716 20818 20833 ACTGAGTGTGCAGTTC  64  967 881387 2829 2844 20946 20961 AAGCAAGTGGAGGTCT  49  968 881411 2913 2928 21030 21045 GTATGAGAAACGGCCT  41  969 881435 3049 3064 21166 21181 GTAATGTAGGGATACA  52  970 881459 3277 3292 21394 21409 GAAGAATAAACACCCC  48  971 881483 3430 3445 21547 21562 TGGCAGTGGAGCTCCA  54  972 881507 3517 3532 21634 21649 CATCACTGCAGTTCAC  53  973 881530 3631 3646 21748 21763 CTTCAGCAAAGAGGGT  41  974 881554 3918 3933 22035 22050 CATAGCTAGTTCATTC  65  975 881578 4089 4104 22206 22221 TCCATATGACCCAGTG  34  976 881597 4305 4320 22422 22437 CAATAGTCTAGCTGGG  37  977 881621 4418 4433 22535 22550 CAAACACTAATACCTA  69  978 881645 4556 4571 22673 22688 GTAACTGACACGCACA  49  979 881669 4618 4633 22735 22750 GGGCATGTGACTGGAG  50  980 881693 4845 4860 22962 22977 GTAACAGGCTATCAGG  55  981 881717 5152 5167 23269 23284 GCAAAACGCAGAGCCA  43  982 881741 5209 5224 23326 23341 ACGACAGCATAGCATT  33  983 881765 N/A N/A 18783 18798 TATATGACGACTTAAG  70  984 881789 N/A N/A 18853 18868 TCCTGAATTTACAGCA  52  985 881834 N/A N/A  4404  4419 CGGGAGAGCGGAGGCG 100  986 881857 N/A N/A  4740  4755 CCGCACTCACTCGCAG  92  987 881881 N/A N/A  5057  5072 CCACGAGGCACCGCAC 119  988 881905 N/A N/A  5668  5683 ACGAGAACGCACGGAC  60  989 881927 N/A N/A  5779  5794 AGACGAAAACAGCCGC  75  990 881949 N/A N/A  6209  6224 ACTAAGGACAGCTGTG  85  991 881972 N/A N/A  6424  6439 GAAAGAATGGCGCATA  51  992 881995 N/A N/A  7168  7183 TTCAACTTGTGACCCA  51  993 882019 N/A N/A  7385  7400 TGCATAGTGATGAGAT  63  994 882043 N/A N/A  7726  7741 TAACACGGTGTTGACA  65  995 882067 N/A N/A  8147  8162 TCCAACAGGCGGACAC  71  996 882090 N/A N/A  8399  8414 GATCATAAAGGACAAT  70  997 882114 N/A N/A  8570  8585 AAGGAGTGCAGGAGAG  87  998  8622  8637  8674  8689  8726  8741  8778  8793  8830  8845  8882  8897  8934  8949  8986  9001  9038  9053  9090  9105 882137 N/A N/A  9387  9402 GAGAAAACCCCCCTCT  83  999 882160 N/A N/A  9611  9626 ACACACCACTATTCGA  65 1000 882184 N/A N/A  9990 10005 CAAGGATAACCTGAAC  74 1001 882207 N/A N/A 10179 10194 GCAGTAAACGCAAGTC  46 1002 882231 N/A N/A 10357 10372 CTGAAGACAATGTTGT  45 1003 882255 N/A N/A 10672 10687 TAGCAGGGCACGCTCT  69 1004 882279 N/A N/A 11090 11105 TCCTAAATCACCCTGG  79 1005 882303 N/A N/A 11370 11385 ACCAGAACTAATGTCC  68 1006 882327 N/A N/A 11674 11689 GGCAAGATGAGAATTC  75 1007 882351 N/A N/A 11987 12002 GAGATATAAATAGCTG  49 1008 882375 N/A N/A 12175 12190 AACTATCTTATTCCTC  70 1009 882422 N/A N/A 12897 12912 AAAAATATAACGGTGT  76 1010 882445 N/A N/A 13203 13218 AACACTTGGAGAAGTC  57 1011 882467 N/A N/A 13904 13919 AGTCAAGCCCCAAGCC  87 1012 882489 N/A N/A 14161 14176 GCATGAGGGTGACACA  58 1013 882512 N/A N/A 14432 14447 TCCCACGCGGGAGGCT  99 1014 882535 N/A N/A 14717 14732 GGACATATTTGCTTTG  68 1015 882559 N/A N/A 15501 15516 TTCCAGCTTAGTCACC  63 1016 882582 N/A N/A 15806 15821 CCTAGATAACATGAGA  74 1017 882605 N/A N/A 15927 15942 CCCAAATGACTTAGTC  68 1018 882629 N/A N/A 16178 16193 ACAATAAAATGTCACG  71 1019 882652 N/A N/A 16394 16409 CTCCAGGGATGGAACA  81 1020 882676 N/A N/A 16705 16720 GACCACAGTGAAGTCG  86 1021 882700 N/A N/A 16926 16941 GCTTACTGTGATTCTG  66 1022 882724 N/A N/A 17241 17256 AAGGGTTACGCTTATT  48 1023 882748 N/A N/A 17545 17560 ATGGATAGTTTCTCAT  75 1024 882772 N/A N/A 17760 17775 TCCTAACCCTACCCTT  70 1025 882796 N/A N/A 17991 18006 GAAATAGCATCAGAGC  55 1026 882819 N/A N/A 18601 18616 ATCCATGTCAACTTTA  41 1027 882843 N/A N/A 18965 18980 CAAAACTACTATTGTG  77 1028 882866 N/A N/A 19241 19256 AGCCAAAGTGACTTAA  42 1029 882892 N/A N/A  4089  4104 CGACAGTCCGGTTAGC  89 1030 882904 N/A N/A 12597 12612 TTCCACACTGGATATG  59 1031

TABLE 16 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  39  195 881078   14   29  3753  3768 ATCGAGCGGTGAAACT  71 1032 881102  357  372  6836  6851 TCGGAACTTTCCTTTA  67 1033 881125  692  707  9182  9197 CATTGGTACGGGATTT  84 1034 881172 1208 1223 13761 13776 GACAAGAACTGCTGTG  85 1035 881196 1444 1459 19561 19576 GGATAGATCTGTGGTA  51 1036 881220 1745 1760 19862 19877 CAAAACAGTAAGAGGG  69 1037 881244 1950 1965 20067 20082 GCGCATTTCAGTAAAT  67 1038 881268 2046 2061 20163 20178 CATACCTACAAGCAGT  57 1039 881292 2199 2214 20316 20331 CAAGGCTACAGGCACG  49 1040 881316 2389 2404 20506 20521 ACATTACTAGATTGTA  59 1041 881340 2525 2540 20642 20657 TTGGACAGAAGGGCAA  71 1042 881364 2710 2725 20827 20842 AAACAGCCCACTGAGT  89 1043 881388 2830 2845 20947 20962 TAAGCAAGTGGAGGTC  53 1044 881412 2914 2929 21031 21046 AGTATGAGAAACGGCC  38 1045 881436 3050 3065 21167 21182 TGTAATGTAGGGATAC  47 1046 881460 3343 3358 21460 21475 GCAAGATGACTCCATG  37 1047 881484 3431 3446 21548 21563 ATGGCAGTGGAGCTCC  44 1048 881508 3520 3535 21637 21652 ATACATCACTGCAGTT  64 1049 881531 3643 3658 21760 21775 GGGTTAAATATTCTTC  48 1050 881555 3919 3934 22036 22051 TCATAGCTAGTTCATT  48 1051 881579 4125 4140 22242 22257 AACCAAGAAATGCACC  67 1052 881598 4306 4321 22423 22438 CCAATAGTCTAGCTGG  81 1053 881622 4419 4434 22536 22551 TCAAACACTAATACCT  75 1054 881646 4557 4572 22674 22689 AGTAACTGACACGCAC  49 1055 881670 4667 4682 22784 22799 TTAACCAATCCCAACA  70 1056 881694 4846 4861 22963 22978 TGTAACAGGCTATCAG  69 1057 881718 5153 5168 23270 23285 AGCAAAACGCAGAGCC  39 1058 881742 5210 5225 23327 23342 AACGACAGCATAGCAT  26 1059 881766 N/A N/A 18784 18799 ATATATGACGACTTAA  94 1060 881790 N/A N/A 18871 18886 GCCTTAATGTTCAGCT  51 1061 881811 N/A N/A  4126  4141 GCCTTGGACGGCCCCG  77 1062 881835 N/A N/A  4416  4431 CCGGAGCAGGCCCGGG 113 1063 881858 N/A N/A  4806  4821 CCGACACGCGCCGCTC  70 1064 881882 N/A N/A  5059  5074 AGCCACGAGGCACCGC  88 1065 881906 N/A N/A  5672  5687 GGAAACGAGAACGCAC  70 1066 881928 N/A N/A  5783  5798 TGAGAGACGAAAACAG 109 1067 881950 N/A N/A  6216  6231 GCCTAGGACTAAGGAC  67 1068 881973 N/A N/A  6426  6441 AAGAAAGAATGGCGCA  37 1069 881996 N/A N/A  7187  7202 TGCTGAACCCCACAGG  67 1070 882020 N/A N/A  7386  7401 ATGCATAGTGATGAGA  55 1071 882044 N/A N/A  7728  7743 CATAACACGGTGTTGA  64 1072 882068 N/A N/A  8148  8163 TTCCAACAGGCGGACA  57 1073 882091 N/A N/A  8406  8421 CATGGAGGATCATAAA  78 1074 882115 N/A N/A  8598  8613 AGGAAGCACTGGCATC  43 1075  8650  8665  9014  9029 882138 N/A N/A  9424  9439 TAAACTTGGCTGTGGG  73 1076 882161 N/A N/A  9632  9647 TTCCAACAATAGCAAC  54 1077 882208 N/A N/A 10181 10196 GAGCAGTAAACGCAAG  46 1078 882232 N/A N/A 10367 10382 AACCATGTTCCTGAAG  59 1079 882256 N/A N/A 10674 10689 ACTAGCAGGGCACGCT  62 1080 882280 N/A N/A 11101 11116 TCTAATGGTGCTCCTA  49 1081 882304 N/A N/A 11378 11393 GATGAGGGACCAGAAC  80 1082 882328 N/A N/A 11675 11690 GGGCAAGATGAGAATT 100 1083 882352 N/A N/A 11989 12004 CCGAGATATAAATAGC  39 1084 882376 N/A N/A 12206 12221 TATCATGCATACCAAA  42 1085 882399 N/A N/A 12654 12669 TGGTAGAATGTGATAT  57 1086 882423 N/A N/A 12900 12915 GCAAAAAATATAACGG  53 1087 882446 N/A N/A 13275 13290 CGTCAAGGAGGCCTGG  57 1088 882468 N/A N/A 13913 13928 CTCTACTGGAGTCAAG  75 1089 882490 N/A N/A 14162 14177 TGCATGAGGGTGACAC  55 1090 882513 N/A N/A 14458 14473 GGCGAGTGGCGGGTAG  92 1091 882536 N/A N/A 14736 14751 CTGAAGCTTAGTTATC  62 1092 882560 N/A N/A 15515 15530 ACTCAATGTGCACCTT  71 1093 882583 N/A N/A 15807 15822 CCCTAGATAACATGAG  83 1094 882606 N/A N/A 15982 15997 ACTTACAGGACTATTT  84 1095 882630 N/A N/A 16200 16215 GACAAATCTTCTGCCT  74 1096 882653 N/A N/A 16422 16437 TGTAACTCTGAGTAGA  57 1097 882677 N/A N/A 16708 16723 GTAGACCACAGTGAAG  71 1098 882701 N/A N/A 16934 16949 AAGCAAGAGCTTACTG  86 1099 882725 N/A N/A 17246 17261 AGTTTAAGGGTTACGC  37 1100 882749 N/A N/A 17546 17561 AATGGATAGTTTCTCA  37 1101 882773 N/A N/A 17771 17786 TCTAACCCTAATCCTA  95 1102 882797 N/A N/A 18030 18045 GTTCAAGATTAAACCA  29 1103 882820 N/A N/A 18605 18620 TTGCATCCATGTCAAC  53 1104 882844 N/A N/A 19017 19032 GTATAGTTCTCAACCA  53 1105 882867 N/A N/A 19266 19281 TCCATAGATCAACATG  55 1106 882903 N/A N/A 9991 10006 CCAAGGATAACCTGAA  56 1107

TABLE 17 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  37  195 881079   15   30  3754  3769 GATCGAGCGGTGAAAC  77 1108 881103  412  427  6891  6906 AAGCGCACCGCAGGCG  96 1109 881126  694  709  9184  9199 GACATTGGTACGGGAT  64 1110 881173 1209 1224 13762 13777 TGACAAGAACTGCTGT  82 1111 881197 1446 1461 19563 19578 GCGGATAGATCTGTGG  45 1112 881221 1777 1792 19894 19909 CGCTGAACTGAAATCT  63 1113 881245 1955 1970 20072 20087 AAAGAGCGCATTTCAG  94 1114 881269 2047 2062 20164 20179 ACATACCTACAAGCAG  55 1115 881293 2202 2217 20319 20334 CCCCAAGGCTACAGGC  75 1116 881317 2391 2406 20508 20523 AGACATTACTAGATTG  34 1117 881341 2530 2545 20647 20662 AGTACTTGGACAGAAG  57 1118 881365 2740 2755 20857 20872 GAGGAAGCATAGAACA  82 1119 881389 2832 2847 20949 20964 CTTAAGCAAGTGGAGG  30 1120 881413 2915 2930 21032 21047 TAGTATGAGAAACGGC  22 1121 881437 3051 3066 21168 21183 GTGTAATGTAGGGATA  36 1122 881461 3344 3359 21461 21476 TGCAAGATGACTCCAT  43 1123 881485 3433 3448 21550 21565 ACATGGCAGTGGAGCT  36 1124 881509 3522 3537 21639 21654 GTATACATCACTGCAG  56 1125 881532 3660 3675 21777 21792 CTTTGAAGTGCTGTGT  84 1126 881556 3920 3935 22037 22052 TTCATAGCTAGTTCAT  56 1127 881580 4168 4183 22285 22300 TTGAGGTTTTCCTAAA  71 1128 881599 4307 4322 22424 22439 CCCAATAGTCTAGCTG  44 1129 881623 4430 4445 22547 22562 GACCAGCTTTTTCAAA  62 1130 881647 4558 4573 22675 22690 AAGTAACTGACACGCA  77 1131 881671 4670 4685 22787 22802 ACCTTAACCAATCCCA  29 1132 881695 4847 4862 22964 22979 CTGTAACAGGCTATCA  56 1133 881719 5154 5169 23271 23286 CAGCAAAACGCAGAGC  46 1134 881743 5212 5227 23329 23344 CCAACGACAGCATAGC  24 1135 881767 N/A N/A 18785 18800 TATATATGACGACTTA  80 1136 881791 N/A N/A 18872 18887 CGCCTTAATGTTCAGC  34 1137 881812 N/A N/A  4152  4167 AGGCAGTTGTGCCGTC 162 1138 881836 N/A N/A  4423  4438 CCGGACCCCGGAGCAG 124 1139 881859 N/A N/A  4807  4822 CCCGACACGCGCCGCT  88 1140 881883 N/A N/A  5062  5077 TTCAGCCACGAGGCAC  95 1141 881907 N/A N/A  5674  5689 GTGGAAACGAGAACGC  97 1142 881929 N/A N/A  5794  5809 CAGAGACGCGGTGAGA 161 1143 881951 N/A N/A  6217  6232 TGCCTAGGACTAAGGA  74 1144 881974 N/A N/A  6479  6494 GCTAAACCCAAAATAC  96 1145 881997 N/A N/A  7192  7207 TCCTATGCTGAACCCC  64 1146 882021 N/A N/A  7390  7405 TACCATGCATAGTGAT  56 1147 882045 N/A N/A  7730  7745 TGCATAACACGGTGTT  62 1148 882069 N/A N/A  8153  8168 GCATATTCCAACAGGC  34 1149 882092 N/A N/A  8409  8424 ACTCATGGAGGATCAT  81 1150 882116 N/A N/A  8599  8614 TAGGAAGCACTGGCAT  72 1151  8651  8666  8755  8770  8859  8874  8911  8926  9015  9030 882139 N/A N/A  9425  9440 GTAAACTTGGCTGTGG  82 1152 882162 N/A N/A  9669  9684 TGGTATTTTTCCGTTC  31 1153 882185 N/A N/A  9992 10007 GCCAAGGATAACCTGA  31 1154 882209 N/A N/A 10186 10201 AGCCAGAGCAGTAAAC  85 1155 882233 N/A N/A 10371 10386 ACTGAACCATGTTCCT  49 1156 882257 N/A N/A 10676 10691 CAACTAGCAGGGCACG  62 1157 882281 N/A N/A 11102 11117 TTCTAATGGTGCTCCT  93 1158 882305 N/A N/A 11396 11411 GCACATCAATGTTTTA  33 1159 882329 N/A N/A 11687 11702 AAAGATGCCAGAGGGC  77 1160 882353 N/A N/A 11991 12006 TGCCGAGATATAAATA  77 1161 882377 N/A N/A 12207 12222 GTATCATGCATACCAA  35 1162 882400 N/A N/A 12675 12690 GCCTTAATGGTGATTT  74 1163 882447 N/A N/A 13289 13304 ACAAAAGGTTCCCGCG  97 1164 882469 N/A N/A 13921 13936 CAGAAGATCTCTACTG  88 1165 882491 N/A N/A 14163 14178 CTGCATGAGGGTGACA  81 1166 882514 N/A N/A 14477 14492 GAAGAGTTGGCGGTGG  85 1167 882537 N/A N/A 14737 14752 CCTGAAGCTTAGTTAT  65 1168 882561 N/A N/A 15531 15546 ACGCAGTGCACCTGTG 107 1169 882584 N/A N/A 15813 15828 CTCCAACCCTAGATAA  98 1170 882607 N/A N/A 15983 15998 AACTTACAGGACTATT  87 1171 882631 N/A N/A 16201 16216 AGACAAATCTTCTGCC 104 1172 882654 N/A N/A 16430 16445 CTGAACTGTGTAACTC  72 1173 882678 N/A N/A 16710 16725 TAGTAGACCACAGTGA  91 1174 882702 N/A N/A 16950 16965 CACCAGGTTAAACAGG 108 1175 882726 N/A N/A 17247 17262 TAGTTTAAGGGTTACG  51 1176 882750 N/A N/A 17554 17569 CTCCAGGGAATGGATA  73 1177 882774 N/A N/A 17777 17792 GAAAACTCTAACCCTA  91 1178 882798 N/A N/A 18087 18102 TTATATACTGGTTGGT  51 1179 882821 N/A N/A 18620 18635 TTAGAGGACAGTGACT  82 1180 882845 N/A N/A 19018 19033 TGTATAGTTCTCAACC  57 1181 882868 N/A N/A 19267 19282 TTCCATAGATCAACAT  56 1182 882905 N/A N/A 12921 12936 CCCTAAGTTTAATTTA 103 1183

TABLE 18 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  38  195 881080   16   31  3755  3770 AGATCGAGCGGTGAAA  76 1184 881104  415  430  6894  6909 TCAAAGCGCACCGCAG  57 1185 881127  697  712  9187  9202 TGGGACATTGGTACGG  31 1186 881174 1210 1225 13763 13778 CTGACAAGAACTGCTG  79 1187 881198 1447 1462 19564 19579 GGCGGATAGATCTGTG  49 1188 881222 1780 1795 19897 19912 AACCGCTGAACTGAAA  87 1189 881246 1957 1972 20074 20089 TTAAAGAGCGCATTTC  59 1190 881270 2048 2063 20165 20180 GACATACCTACAAGCA  50 1191 881294 2229 2244 20346 20361 AACCGCTGGCAGGTGG  60 1192 881318 2393 2408 20510 20525 TTAGACATTACTAGAT  55 1193 881342 2531 2546 20648 20663 AAGTACTTGGACAGAA  46 1194 881366 2743 2758 20860 20875 CACGAGGAAGCATAGA  44 1195 881390 2833 2848 20950 20965 ACTTAAGCAAGTGGAG  37 1196 881414 2917 2932 21034 21049 TGTAGTATGAGAAACG  31 1197 881438 3059 3074 21176 21191 GCTGAACTGTGTAATG  42 1198 881462 3345 3360 21462 21477 GTGCAAGATGACTCCA  44 1199 881486 3435 3450 21552 21567 GTACATGGCAGTGGAG  44 1200 881510 3523 3538 21640 21655 TGTATACATCACTGCA  37 1201 881533 3665 3680 21782 21797 AGCTTCTTTGAAGTGC  38 1202 881557 3921 3936 22038 22053 TTTCATAGCTAGTTCA  44 1203 881581 4170 4185 22287 22302 GCTTGAGGTTTTCCTA  23 1204 881600 4312 4327 22429 22444 TCATACCCAATAGTCT  48 1205 881624 4438 4453 22555 22570 CGCTCAAAGACCAGCT  44 1206 881648 4559 4574 22676 22691 AAAGTAACTGACACGC  33 1207 881672 4671 4686 22788 22803 GACCTTAACCAATCCC  36 1208 881696 4863 4878 22980 22995 GACCTTTACTTCATTC  48 1209 881720 5157 5172 23274 23289 TCTCAGCAAAACGCAG  73 1210 881744 5213 5228 23330 23345 ACCAACGACAGCATAG  33 1211 881768 N/A N/A 18786 18801 TTATATATGACGACTT  65 1212 881813 N/A N/A  4156  4171 TCGCAGGCAGTTGTGC 120 1213 881837 N/A N/A  4425  4440 CGCCGGACCCCGGAGC  91 1214 881860 N/A N/A  4816  4831 CCAAAGGCTCCCGACA  71 1215 881884 N/A N/A  5065  5080 CCCTTCAGCCACGAGG  92 1216 881908 N/A N/A  5675  5690 CGTGGAAACGAGAACG  71 1217 881952 N/A N/A  6229  6244 TCTGAGTGAGCTTGCC  51 1218 881975 N/A N/A  6541  6556 TGCATAGGCATCCTTC  32 1219 881998 N/A N/A  7200  7215 ATTAATTCTCCTATGC  85 1220 882022 N/A N/A  7393  7408 TATTACCATGCATAGT  70 1221 882046 N/A N/A  7732  7747 AATGCATAACACGGTG  51 1222 882070 N/A N/A  8154  8169 AGCATATTCCAACAGG  31 1223 882093 N/A N/A  8417  8432 GTGAAAACACTCATGG  40 1224 882117 N/A N/A  8606  8621 GCTGAGATAGGAAGCA  81 1225  8658  8673  8762  8777  8866  8881  8918  8933  9022  9037 882140 N/A N/A  9426  9441 AGTAAACTTGGCTGTG  62 1226 882163 N/A N/A  9710  9725 TGGTAGGTAGGCACCT  65 1227 882186 N/A N/A 10006 10021 CTTCACTACCTGCAGC  78 1228 882210 N/A N/A 10190 10205 ATAGAGCCAGAGCAGT  34 1229 882234 N/A N/A 10372 10387 CACTGAACCATGTTCC  39 1230 882258 N/A N/A 10677 10692 GCAACTAGCAGGGCAC  47 1231 882282 N/A N/A 11116 11131 AGTGATGTCAGGTTTT  28 1232 882306 N/A N/A 11398 11413 AGGCACATCAATGTTT  49 1233 882330 N/A N/A 11688 11703 TAAAGATGCCAGAGGG  61 1234 882354 N/A N/A 11999 12014 CACGAGGTTGCCGAGA  36 1235 882378 N/A N/A 12209 12224 CTGTATCATGCATACC  39 1236 882401 N/A N/A 12682 12697 CACTGAGGCCTTAATG  66 1237 882424 N/A N/A 12927 12942 AATGAACCCTAAGTTT  97 1238 882448 N/A N/A 13290 13305 AACAAAAGGTTCCCGC  60 1239 882470 N/A N/A 13922 13937 ACAGAAGATCTCTACT  68 1240 882492 N/A N/A 14205 14220 GTACAGTCCACTCCAC  90 1241 882515 N/A N/A 14550 14565 GGCACATGAGAAATCA  67 1242 882538 N/A N/A 14993 15008 AACTGCAGCACCGTGG  55 1243 882562 N/A N/A 15533 15548 TCACGCAGTGCACCTG  54 1244 882585 N/A N/A 15826 15841 TGAGAAAGCATGGCTC  60 1245 882608 N/A N/A 15987 16002 AGCTAACTTACAGGAC 104 1246 882632 N/A N/A 16208 16223 GTACACAAGACAAATC  79 1247 882655 N/A N/A 16433 16448 TGCCTGAACTGTGTAA  65 1248 882679 N/A N/A 16713 16728 AGGTAGTAGACCACAG  41 1249 882703 N/A N/A 16955 16970 GAACACACCAGGTTAA 102 1250 882727 N/A N/A 17248 17263 CTAGTTTAAGGGTTAC  73 1251 882751 N/A N/A 17565 17580 CTACAAAATGCCTCCA  58 1252 882775 N/A N/A 17778 17793 AGAAAACTCTAACCCT  75 1253 882799 N/A N/A 18089 18104 GATTATATACTGGTTG  41 1254 882822 N/A N/A 18623 18638 GACTTAGAGGACAGTG  52 1255 882846 N/A N/A 19019 19034 CTGTATAGTTCTCAAC  48 1256 882869 N/A N/A 19283 19298 ATTTTTTGGTTAGTCC  47 1257 882896 N/A N/A 5796 5811 AACAGAGACGCGGTGA  90 1258

TABLE 19 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  38  195 881081   18   33  3757  3772 CAAGATCGAGCGGTGA  62 1259 881105  416  431  6895  6910 TTCAAAGCGCACCGCA  51 1260 881128  728  743  9218  9233 TGCCAGTGGTGGCCGC  70 1261 881175 1211 1226 N/A N/A TCTGACAAGAACTGCT  88 1262 881199 1448 1463 19565 19580 TGGCGGATAGATCTGT  66 1263 881223 1784 1799 19901 19916 CCTCAACCGCTGAACT  70 1264 881247 1958 1973 20075 20090 ATTAAAGAGCGCATTT  87 1265 881271 2049 2064 20166 20181 AGACATACCTACAAGC  50 1266 881295 2233 2248 20350 20365 AGGAAACCGCTGGCAG  44 1267 881319 2395 2410 20512 20527 ACTTAGACATTACTAG  80 1268 881343 2534 2549 20651 20666 GTTAAGTACTTGGACA  68 1269 881367 2745 2760 20862 20877 GGCACGAGGAAGCATA  38 1270 881391 2835 2850 20952 20967 ATACTTAAGCAAGTGG  24 1271 881415 2918 2933 21035 21050 CTGTAGTATGAGAAAC  55 1272 881439 3066 3081 21183 21198 GATAAAGGCTGAACTG  33 1273 881463 3359 3374 21476 21491 CTGCATGAAAGTGTGT  44 1274 881487 3443 3458 21560 21575 CCTACTGGGTACATGG  37 1275 881511 3524 3539 21641 21656 ATGTATACATCACTGC  33 1276 881534 3686 3701 21803 21818 CCTGAGACAGACTTCC  45 1277 881558 3924 3939 22041 22056 GTATTTCATAGCTAGT  31 1278 881582 4172 4187 22289 22304 CTGCTTGAGGTTTTCC  26 1279 881601 4314 4329 22431 22446 GTTCATACCCAATAGT  34 1280 881625 4449 4464 22566 22581 ATTTATGCCCTCGCTC  74 1281 881649 4560 4575 22677 22692 AAAAGTAACTGACACG  39 1282 881673 4675 4690 22792 22807 TAAAGACCTTAACCAA  70 1283 881697 4909 4924 23026 23041 TTATGCATGTTCCTCA  43 1284 881721 5164 5179 23281 23296 CTACAGATCTCAGCAA  82 1285 881745 5216 5231 23333 23348 TATACCAACGACAGCA  51 1286 881769 N/A N/A 18787 18802 TTTATATATGACGACT  61 1287 881793 N/A N/A 19455 19470 TAGCAGAGGTTCTACC  74 1288 881814 N/A N/A  4215  4230 AGGAACGAAGAGCAGG  57 1289 881838 N/A N/A  4447  4462 GGGATTCCGCGCGCAG  86 1290 881861 N/A N/A  4818  4833 GCCCAAAGGCTCCCGA  89 1291 881885 N/A N/A  5362  5377 CCGGAGACCTTGAAGA  91 1292 881930 N/A N/A  5797  5812 AAACAGAGACGCGGTG  82 1293 881953 N/A N/A  6241  6256 ACCCACCCAGTGTCTG  82 1294 881976 N/A N/A  6542  6557 TTGCATAGGCATCCTT  53 1295 881999 N/A N/A  7203  7218 GTGATTAATTCTCCTA  30 1296 882023 N/A N/A  7414  7429 CACTTTAAAAGTTGAC  72 1297 882047 N/A N/A  7733  7748 GAATGCATAACACGGT  39 1298 882071 N/A N/A  8158  8173 GAGAAGCATATTCCAA  43 1299 882094 N/A N/A  8431  8446 CTTCATCAGACTAAGT  69 1300 882118 N/A N/A  8607  8622 GGCTGAGATAGGAAGC  64 1301  8659  8674  8763  8778  8867  8882  8919  8934  9023  9038 882141 N/A N/A  9427  9442 GAGTAAACTTGGCTGT  62 1302 882164 N/A N/A  9716  9731 ACCCAGTGGTAGGTAG  70 1303 882187 N/A N/A 10014 10029 GTACACCTCTTCACTA  93 1304 882211 N/A N/A 10191 10206 CATAGAGCCAGAGCAG  40 1305 882235 N/A N/A 10448 10463 GCCAAAGAGCCCAATT  67 1306 882259 N/A N/A 10772 10787 GTCCGACGCACCGCGG  82 1307 882283 N/A N/A 11122 11137 ACAAGCAGTGATGTCA  27 1308 882307 N/A N/A 11403 11418 CTCTTAGGCACATCAA  39 1309 882331 N/A N/A 11689 11704 TTAAAGATGCCAGAGG  72 1310 882355 N/A N/A 12000 12015 TCACGAGGTTGCCGAG  38 1311 882379 N/A N/A 12221 12236 CTGATAATTAATCTGT  37 1312 882402 N/A N/A 12689 12704 GGGTAAGCACTGAGGC  39 1313 882425 N/A N/A 12990 13005 TTTAAGTCATGTGTCA  50 1314 882449 N/A N/A 13291 13306 GAACAAAAGGTTCCCG  82 1315 882471 N/A N/A 13923 13938 GACAGAAGATCTCTAC  83 1316 882493 N/A N/A 14210 14225 AACCAGTACAGTCCAC  65 1317 882516 N/A N/A 14551 14566 AGGCACATGAGAAATC  77 1318 882539 N/A N/A 15020 15035 GGCAATGGAGTCTCGC  50 1319 882563 N/A N/A 15535 15550 TCTCACGCAGTGCACC  56 1320 882586 N/A N/A 15837 15852 GCACAATTCTCTGAGA  66 1321 882609 N/A N/A 16031 16046 TATCAAAGATCTCCAC  75 1322 882633 N/A N/A 16231 16246 CTTTAGCCCATGCTCC  80 1323 882656 N/A N/A 16438 16453 CACTATGCCTGAACTG  90 1324 882680 N/A N/A 16718 16733 TCAAAAGGTAGTAGAC  71 1325 882704 N/A N/A 16959 16974 CACCGAACACACCAGG  80 1326 882728 N/A N/A 17287 17302 TCAGACTGTGCTGCTC  64 1327 882752 N/A N/A 17566 17581 CCTACAAAATGCCTCC  53 1328 882776 N/A N/A 17808 17823 GTTATCTATGGAAACC  58 1329 882800 N/A N/A 18090 18105 GGATTATATACTGGTT  23 1330 882823 N/A N/A 18682 18697 ATGGACCACGCAGCCT  61 1331 882847 N/A N/A 19026 19041 AGGTAATCTGTATAGT  38 1332 882870 N/A N/A 19311 19326 TTCATATTTGGAGCCA  34 1333 882894 N/A N/A  5700  5715 GCCCGGAGGAAGGGCG 106 1334

TABLE 20 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages Compound SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID: 2 IRF4 SEQ Number Start Site Stop Site Start Site Stop Site Sequence (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  26  195 881082   20   35  3759  3774 CCCAAGATCGAGCGGT  72 1335 881106  417  432  6896  6911 GTTCAAAGCGCACCGC  35 1336 881129  741  756  9231  9246 ACAAGCTGGGCCTTGC  73 1337 881152  942  957 13495 13510 GGAGATCCGGCAGCCC  77 1338 881176 1222 1237 N/A N/A ACGCTTGCAGCTCTGA  52 1339 881200 1452 1467 19569 19584 GGAATGGCGGATAGAT  67 1340 881224 1785 1800 19902 19917 TCCTCAACCGCTGAAC  63 1341 881248 1959 1974 20076 20091 AATTAAAGAGCGCATT  83 1342 881272 2050 2065 20167 20182 CAGACATACCTACAAG  56 1343 881296 2234 2249 20351 20366 CAGGAAACCGCTGGCA  26 1344 881320 2396 2411 20513 20528 TACTTAGACATTACTA  61 1345 881344 2536 2551 20653 20668 TAGTTAAGTACTTGGA  44 1346 881368 2746 2761 20863 20878 TGGCACGAGGAAGCAT  54 1347 881392 2836 2851 20953 20968 TATACTTAAGCAAGTG  53 1348 881416 2919 2934 21036 21051 CCTGTAGTATGAGAAA  43 1349 881440 3068 3083 21185 21200 TTGATAAAGGCTGAAC  33 1350 881464 3364 3379 21481 21496 GAGCACTGCATGAAAG  49 1351 881488 3444 3459 21561 21576 CCCTACTGGGTACATG  63 1352 881512 3525 3540 21642 21657 GATGTATACATCACTG  30 1353 881535 3722 3737 21839 21854 GAGCATCCGCTTGGAG  69 1354 881559 3926 3941 22043 22058 GAGTATTTCATAGCTA  29 1355 881583 4173 4188 22290 22305 ACTGCTTGAGGTTTTC  42 1356 881602 4319 4334 22436 22451 TTTTAGTTCATACCCA  48 1357 881626 4450 4465 22567 22582 TATTTATGCCCTCGCT  74 1358 881650 4576 4591 22693 22708 TCTGAATTGTTACTAA  65 1359 881674 4677 4692 22794 22809 AATAAAGACCTTAACC  89 1360 881698 4911 4926 23028 23043 TCTTATGCATGTTCCT  38 1361 881722 5176 5191 23293 23308 AAGCATCCTTTCCTAC  65 1362 881746 5217 5232 23334 23349 GTATACCAACGACAGC  22 1363 881770 N/A N/A 18788 18803 TTTTATATATGACGAC  66 1364 881794 N/A N/A 22820 22835 GAGGACTTCGGGAGAT  77 1365 881815 N/A N/A  4217  4232 TTAGGAACGAAGAGCA  68 1366 881839 N/A N/A  4487  4502 GACAAGTGGCGCAGAC  89 1367 881862 N/A N/A  4853  4868 CTCCTACCCGCCTGCT  85 1368 881886 N/A N/A  5364  5379 GGCCGGAGACCTTGAA  97 1369 881909 N/A N/A  5712  5727 CGGCAGACGGGAGCCC  88 1370 881931 N/A N/A  5798  5813 GAAACAGAGACGCGGT  99 1371 881954 N/A N/A  6265  6280 GCGGAGGTTCCTTGAG  56 1372 882000 N/A N/A  7211  7226 CATACAGTGTGATTAA  47 1373 882024 N/A N/A  7424  7439 GACCATGTTACACTTT  50 1374 882048 N/A N/A  7736  7751 TTAGAATGCATAACAC  56 1375 882072 N/A N/A  8159  8174 TGAGAAGCATATTCCA  28 1376 882095 N/A N/A  8443  8458 CTGGAGTGAACCCTTC  44 1377 882119 N/A N/A  8701  8716 AGAAGCACTGGCATCG  63 1378  8805  8820  8961  8976 882142 N/A N/A  9430  9445 TGAGAGTAAACTTGGC  35 1379 882165 N/A N/A  9742  9757 ATCCACAATCAGCAAG  49 1380 882188 N/A N/A 10016 10031 TGGTACACCTCTTCAC  68 1381 882212 N/A N/A 10192 10207 CCATAGAGCCAGAGCA  36 1382 882236 N/A N/A 10464 10479 GTCTACTTGAGTCTGT  49 1383 882260 N/A N/A 10780 10795 GACAGAGAGTCCGACG 101 1384 882284 N/A N/A 11123 11138 CACAAGCAGTGATGTC  40 1385 882308 N/A N/A 11405 11420 TACTCTTAGGCACATC  41 1386 882332 N/A N/A 11723 11738 TCAGAATTTAGTTAGT  67 1387 882356 N/A N/A 12001 12016 ATCACGAGGTTGCCGA  40 1388 882380 N/A N/A 12223 12238 GGCTGATAATTAATCT  53 1389 882403 N/A N/A 12695 12710 ATTAAAGGGTAAGCAC  68 1390 882426 N/A N/A 13007 13022 TGATAGTGGTGATGTC  46 1391 882450 N/A N/A 13292 13307 GGAACAAAAGGTTCCC  88 1392 882472 N/A N/A 13956 13971 TCTGATCCGGACTCTC  59 1393 882494 N/A N/A 14213 14228 CAAAACCAGTACAGTC  51 1394 882517 N/A N/A 14622 14637 CCAGAGCACACAGACG  64 1395 882540 N/A N/A 15021 15036 GGGCAATGGAGTCTCG  76 1396 882564 N/A N/A 15538 15553 GTCTCTCACGCAGTGC  50 1397 882587 N/A N/A 15838 15853 GGCACAATTCTCTGAG  57 1398 882610 N/A N/A 16033 16048 GATATCAAAGATCTCC  43 1399 882634 N/A N/A 16254 16269 AAAGACAAGTGCCCAT  76 1400 882657 N/A N/A 16441 16456 TGTCACTATGCCTGAA  73 1401 882681 N/A N/A 16726 16741 ACGAAGATTCAAAAGG  64 1402 882705 N/A N/A 16962 16977 CATCACCGAACACACC  76 1403 882729 N/A N/A 17289 17304 CCTCAGACTGTGCTGC  70 1404 882753 N/A N/A 17567 17582 ACCTACAAAATGCCTC  58 1405 882777 N/A N/A 17826 17841 GGCAAATTAATGCTTC  24 1406 882801 N/A N/A 18096 18111 TCTATGGGATTATATA  74 1407 882824 N/A N/A 18684 18699 TTATGGACCACGCAGC  69 1408 882848 N/A N/A 19030 19045 TTCTAGGTAATCTGTA  62 1409 882871 N/A N/A 19312 19327 TTTCATATTTGGAGCC  25 1410 882899 N/A N/A  6633  6648 AGTAGCTGGGCCCTCG  38 1411

TABLE 21 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 31 195 668815 436 451 6915 6930 CCTCAAAGTCATTGCT 79 1412 881083 21 36 3760 3775 TCCCAAGATCGAGCGG 84 1413 881130 743 758 9233 9248 TCACAAGCTGGGCCTT 70 1414 881153 946 961 13499 13514 CATGGGAGATCCGGCA 59 1415 881177 1235 1250 17028 17043 CCGTGGTGAGCAAACG 68 1416 881201 1453 1468 19570 19585 AGGAATGGCGGATAGA 62 1417 881225 1793 1808 19910 19925 CGCAATTCTCCTCAAC 62 1418 881249 1960 1975 20077 20092 AAATTAAAGAGCGCAT 87 1419 881273 2054 2069 20171 20186 GGCACAGACATACCTA 52 1420 881297 2237 2252 20354 20369 CACCAGGAAACCGCTG 85 1421 881321 2399 2414 20516 20531 CATTACTTAGACATTA 81 1422 881345 2537 2552 20654 20669 ATAGTTAAGTACTTGG 52 1423 881369 2752 2767 20869 20884 TATAATTGGCACGAGG 59 1424 881393 2837 2852 20954 20969 GTATACTTAAGCAAGT 58 1425 881417 2921 2936 21038 21053 ATCCTGTAGTATGAGA 55 1426 881441 3069 3084 21186 21201 CTTGATAAAGGCTGAA 40 1427 881465 3372 3387 21489 21504 GCTACAAAGAGCACTG 37 1428 881489 3449 3464 21566 21581 TCAAACCCTACTGGGT 92 1429 881513 3532 3547 21649 21664 TCTATAAGATGTATAC 74 1430 881536 3726 3741 21843 21858 AATGGAGCATCCGCTT 107 1431 881560 3945 3960 22062 22077 TGCTAGGATTCCTAAC 74 1432 881584 4174 4189 22291 22306 TACTGCTTGAGGTTTT 50 1433 881603 4320 4335 22437 22452 TTTTTAGTTCATACCC 59 1434 881627 4451 4466 22568 22583 GTATTTATGCCCTCGC 47 1435 881651 4577 4592 22694 22709 ATCTGAATTGTTACTA 88 1436 881675 4732 4747 22849 22864 CACCATTCAGACAGAT 167 1437 881699 4918 4933 23035 23050 CTACATTTCTTATGCA 61 1438 881723 5180 5195 23297 23312 TGTGAAGCATCCTTTC 55 1439 881747 5218 5233 23335 23350 TGTATACCAACGACAG 42 1440 881771 N/A N/A 18804 18819 GATATTAGAGACTGTA 49 1441 881795 N/A N/A 22823 22838 ACAGAGGACTTCGGGA 136 1442 881816 N/A N/A 4218 4233 CTTAGGAACGAAGAGC 93 1443 881840 N/A N/A 4491 4506 AAACGACAAGTGGCGC 79 1444 881863 N/A N/A 4860 4875 GCGAAGGCTCCTACCC 115 1445 881887 N/A N/A 5369 5384 CCCGAGGCCGGAGACC 94 1446 881910 N/A N/A 5720 5735 GACGGAGGCGGCAGAC 143 1447 881932 N/A N/A 5815 5830 GAAAAGCAGCGAAAAG 98 1448 881955 N/A N/A 6278 6293 GGTAGAGTGAGATGCG 42 1449 881977 N/A N/A 6663 6678 GTTCTAGGGTCCTCCT 50 1450 882001 N/A N/A 7213 7228 GGCATACAGTGTGATT 75 1451 882025 N/A N/A 7428 7443 AGCTGACCATGTTACA 119 1452 882049 N/A N/A 7737 7752 CTTAGAATGCATAACA 74 1453 882073 N/A N/A 8213 8228 GGCACTACTTCCAAAA 69 1454 882096 N/A N/A 8452 8467 CCGAAAAGACTGGAGT 43 1455 882120 N/A N/A 8702 8717 AAGAAGCACTGGCATC 77 1456 8806 8821 8962 8977 882143 N/A N/A 9432 9447 CCTGAGAGTAAACTTG 82 1457 882166 N/A N/A 9745 9760 CTAATCCACAATCAGC 55 1458 882189 N/A N/A 10024 10039 TTAAAGAGTGGTACAC 94 1459 882213 N/A N/A 10194 10209 TTCCATAGAGCCAGAG 63 1460 882237 N/A N/A 10497 10512 CAAAGCGGGTTCACAA 84 1461 882261 N/A N/A 10781 10796 AGACAGAGAGTCCGAC 93 1462 882285 N/A N/A 11126 11141 AACCACAAGCAGTGAT 83 1463 882309 N/A N/A 11409 11424 GTATTACTCTTAGGCA 36 1464 882333 N/A N/A 11750 11765 GCCACAACTCTCGCCT 75 1465 882357 N/A N/A 12004 12019 GAAATCACGAGGTTGC 38 1466 882381 N/A N/A 12241 12256 TCTAATAAATGTGCTC 56 1467 882404 N/A N/A 12698 12713 CACATTAAAGGGTAAG 101 1468 882427 N/A N/A 13049 13064 ACTATTAAGAACTTGC 79 1469 882451 N/A N/A 13293 13308 GGGAACAAAAGGTTCC 111 1470 882473 N/A N/A 13957 13972 ATCTGATCCGGACTCT 91 1471 882495 N/A N/A 14214 14229 GCAAAACCAGTACAGT 42 1472 882518 N/A N/A 14630 14645 CAACAGTGCCAGAGCA 89 1473 882541 N/A N/A 15070 15085 TCCTATGGTCAGCCTC 52 1474 882565 N/A N/A 15590 15605 CGCAAGTCTACAGCCC 41 1475 882588 N/A N/A 15839 15854 TGGCACAATTCTCTGA 87 1476 882611 N/A N/A 16042 16057 GAATAGGAAGATATCA 94 1477 882635 N/A N/A 16256 16271 GGAAAGACAAGTGCCC 46 1478 882658 N/A N/A 16447 16462 TCCCACTGTCACTATG 85 1479 882682 N/A N/A 16727 16742 CACGAAGATTCAAAAG 90 1480 882706 N/A N/A 16971 16986 AGAAACCCTCATCACC 105 1481 882730 N/A N/A 17330 17345 CTTTGATGAGCAGATC 88 1482 882754 N/A N/A 17570 17585 GAGACCTACAAAATGC 89 1483 882778 N/A N/A 17856 17871 GTCTTAAGGGTTCAAG 51 1484 882802 N/A N/A 18097 18112 GTCTATGGGATTATAT 79 1485 882825 N/A N/A 18685 18700 TTTATGGACCACGCAG 62 1486 882849 N/A N/A 19033 19048 GATTTCTAGGTAATCT 69 1487 882872 N/A N/A 19337 19352 TTACATCCTAGAACAC 97 1488

TABLE 22 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 35 195 669124 3537 3552 21654 21669 TTGCATCTATAAGATG 69 1489 881084 44 59 3783 3798 TCGGAGCTGAGGGCAG 82 1490 881107 448 463 6927 6942 GCTCAACCAGTTCCTC 62 1491 881131 754 769 N/A N/A GGCAACCATTTTCACA 117 1492 881154 949 964 13502 13517 GTCCATGGGAGATCCG 48 1493 881178 1245 1260 17038 17053 CAGGGAGCGGCCGTGG 83 1494 881202 1454 1469 19571 19586 GAGGAATGGCGGATAG 69 1495 881226 1794 1809 19911 19926 CCGCAATTCTCCTCAA 80 1496 881250 1961 1976 20078 20093 CAAATTAAAGAGCGCA 43 1497 881274 2055 2070 20172 20187 TGGCACAGACATACCT 62 1498 881298 2240 2255 20357 20372 CCACACCAGGAAACCG 86 1499 881322 2400 2415 20517 20532 CCATTACTTAGACATT 36 1500 881346 2538 2553 20655 20670 GATAGTTAAGTACTTG 51 1501 881370 2756 2771 20873 20888 AAACTATAATTGGCAC 70 1502 881394 2838 2853 20955 20970 GGTATACTTAAGCAAG 32 1503 881418 2927 2942 21044 21059 GTAAATATCCTGTAGT 67 1504 881442 3070 3085 21187 21202 GCTTGATAAAGGCTGA 27 1505 881466 3373 3388 21490 21505 AGCTACAAAGAGCACT 67 1506 881490 3450 3465 21567 21582 GTCAAACCCTACTGGG 69 1507 881537 3729 3744 21846 21861 TGAAATGGAGCATCCG 91 1508 881561 3951 3966 22068 22083 GACAAGTGCTAGGATT 39 1509 881585 4176 4191 22293 22308 ATTACTGCTTGAGGTT 38 1510 881604 4321 4336 22438 22453 CTTTTTAGTTCATACC 63 1511 881628 4452 4467 22569 22584 TGTATTTATGCCCTCG 56 1512 881652 4580 4595 22697 22712 TGGATCTGAATTGTTA 45 1513 881676 4735 4750 22852 22867 ACGCACCATTCAGACA 102 1514 881700 4972 4987 23089 23104 GGAGGAGGGTTGGCCT 99 1515 881724 5181 5196 23298 23313 TTGTGAAGCATCCTTT 37 1516 881748 5219 5234 23336 23351 ATGTATACCAACGACA 37 1517 881772 N/A N/A 18805 18820 TGATATTAGAGACTGT 40 1518 881796 N/A N/A 22825 22840 CCACAGAGGACTTCGG 73 1519 881817 N/A N/A 4220 4235 CCCTTAGGAACGAAGA 106 1520 881841 N/A N/A 4494 4509 TGCAAACGACAAGTGG 79 1521 881864 N/A N/A 4861 4876 CGCGAAGGCTCCTACC 141 1522 881888 N/A N/A 5370 5385 TCCCGAGGCCGGAGAC 127 1523 881911 N/A N/A 5723 5738 ACGGACGGAGGCGGCA 89 1524 881956 N/A N/A 6279 6294 CGGTAGAGTGAGATGC 46 1525 881978 N/A N/A 6677 6692 CATCGAACTCCTGAGT 69 1526 882002 N/A N/A 7218 7233 ATTGAGGCATACAGTG 70 1527 882026 N/A N/A 7437 7452 AAACACCTCAGCTGAC 84 1528 882050 N/A N/A 7784 7799 GACCTAACTAAATGTC 118 1529 882074 N/A N/A 8218 8233 GATAAGGCACTACTTC 53 1530 882097 N/A N/A 8463 8478 CATTTTATCATCCGAA 45 1531 882121 N/A N/A 8753 8768 GGAAGCACTGGCATTG 51 1532 8857 8872 8909 8924 882144 N/A N/A 9442 9457 TAGCATGGATCCTGAG 100 1533 882167 N/A N/A 9746 9761 TCTAATCCACAATCAG 59 1534 882190 N/A N/A 10025 10040 ATTAAAGAGTGGTACA 107 1535 882214 N/A N/A 10198 10213 CTAATTCCATAGAGCC 20 1536 882238 N/A N/A 10500 10515 TCACAAAGCGGGTTCA 77 1537 882262 N/A N/A 10785 10800 GTCTAGACAGAGAGTC 98 1538 882286 N/A N/A 11151 11166 AGCCAGTGGCTGAAAC 79 1539 882310 N/A N/A 11411 11426 GTGTATTACTCTTAGG 14 1540 882334 N/A N/A 11751 11766 TGCCACAACTCTCGCC 69 1541 882358 N/A N/A 12005 12020 AGAAATCACGAGGTTG 36 1542 882382 N/A N/A 12260 12275 TCAATGGGAATCACAG 60 1543 882405 N/A N/A 12699 12714 ACACATTAAAGGGTAA 89 1544 882428 N/A N/A 13050 13065 CACTATTAAGAACTTG 67 1545 882452 N/A N/A 13309 13324 CCCACATGTCCCGTGG 95 1546 882474 N/A N/A 13995 14010 TTGCAAGAGAACAGCC 61 1547 882496 N/A N/A 14215 14230 TGCAAAACCAGTACAG 72 1548 882519 N/A N/A 14639 14654 AGCACATGTCAACAGT 69 1549 882542 N/A N/A 15091 15106 AACCAGCACAGTTCTC 118 1550 882566 N/A N/A 15591 15606 GCGCAAGTCTACAGCC 120 1551 882589 N/A N/A 15854 15869 ATCAGAATGGCGAGTT 53 1552 882612 N/A N/A 16046 16061 ACCTGAATAGGAAGAT 69 1553 882636 N/A N/A 16258 16273 TTGGAAAGACAAGTGC 85 1554 882659 N/A N/A 16464 16479 AGCCATCGGCAGCTGA 86 1555 882683 N/A N/A 16733 16748 TGGACCCACGAAGATT 62 1556 882707 N/A N/A 16972 16987 CAGAAACCCTCATCAC 96 1557 882731 N/A N/A 17367 17382 AAAGAGGCACCCTCCT 138 1558 882755 N/A N/A 17588 17603 TGCTAGGACACAGCTG 57 1559 882779 N/A N/A 17862 17877 TCAAAAGTCTTAAGGG 93 1560 882803 N/A N/A 18111 18126 GAGAAAACTCCTGAGT 95 1561 882826 N/A N/A 18686 18701 TTTTATGGACCACGCA 69 1562 882850 N/A N/A 19079 19094 CACAACTGCAGTTTGA 86 1563 882873 N/A N/A 19343 19358 CCAAAGTTACATCCTA 61 1564 882897 N/A N/A 5887 5902 CCGCAGAGAGCTAGCA 106 1565

TABLE 23 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 48 195 881085 49 64 3788 3803 TGGACTCGGAGCTGAG 76 1566 881108 462 477 6941 6956 GTCCAGCTGGCTCCGC 47 1567 881132 762 777 N/A N/A TGTCACCTGGCAACCA 91 1568 881155 987 1002 13540 13555 GAACAGGACCTGGTCC 75 1569 881203 1455 1470 19572 19587 AGAGGAATGGCGGATA 58 1570 881251 1962 1977 20079 20094 ACAAATTAAAGAGCGC 36 1571 881275 2078 2093 20195 20210 TTACATCTTACTTCCC 58 1572 881299 2306 2321 20423 20438 ACTAACTGGCTTCCAG 69 1573 881323 2401 2416 20518 20533 ACCATTACTTAGACAT 35 1574 881347 2539 2554 20656 20671 AGATAGTTAAGTACTT 89 1575 881371 2758 2773 20875 20890 TCAAACTATAATTGGC 26 1576 881395 2839 2854 20956 20971 AGGTATACTTAAGCAA 32 1577 881419 2929 2944 21046 21061 TAGTAAATATCCTGTA 58 1578 881443 3071 3086 21188 21203 AGCTTGATAAAGGCTG 64 1579 881467 3376 3391 21493 21508 GTTAGCTACAAAGAGC 59 1580 881491 3451 3466 21568 21583 TGTCAAACCCTACTGG 77 1581 881514 3549 3564 21666 21681 CCCCAAAATACTTTGC 0 1582 881538 3730 3745 21847 21862 TTGAAATGGAGCATCC 78 1583 881562 3952 3967 22069 22084 AGACAAGTGCTAGGAT 44 1584 881586 4186 4201 22303 22318 GGAGATATTAATTACT 50 1585 881605 4369 4384 22486 22501 GCTCATTTCACTGTAG 36 1586 881629 4453 4468 22570 22585 CTGTATTTATGCCCTC 40 1587 881653 4584 4599 22701 22716 ACACTGGATCTGAATT 54 1588 881677 4742 4757 22859 22874 AGCCTTCACGCACCAT 70 1589 881701 4976 4991 23093 23108 CATTGGAGGAGGGTTG 102 1590 881725 5183 5198 23300 23315 GTTTGTGAAGCATCCT 35 1591 881749 5220 5235 23337 23352 GATGTATACCAACGAC 32 1592 881773 N/A N/A 18806 18821 ATGATATTAGAGACTG 26 1593 881797 N/A N/A 3804 3819 AGCCCTTACCTCGCCC 75 1594 881818 N/A N/A 4239 4254 GGCCGCCGTGAGCTTG 97 1595 881865 N/A N/A 4862 4877 CCGCGAAGGCTCCTAC 86 1596 881912 N/A N/A 5724 5739 CACGGACGGAGGCGGC 82 1597 881933 N/A N/A 5894 5909 GGAGTACCCGCAGAGA 75 1598 881957 N/A N/A 6282 6297 AACCGGTAGAGTGAGA 94 1599 881979 N/A N/A 6682 6697 CGCAGCATCGAACTCC 67 1600 882003 N/A N/A 7219 7234 CATTGAGGCATACAGT 45 1601 882027 N/A N/A 7439 7454 ATAAACACCTCAGCTG 79 1602 882051 N/A N/A 7791 7806 AGGAACTGACCTAACT 69 1603 882075 N/A N/A 8219 8234 TGATAAGGCACTACTT 65 1604 882098 N/A N/A 8464 8479 GCATTTTATCATCCGA 37 1605 882122 N/A N/A 8754 8769 AGGAAGCACTGGCATT 41 1606 8858 8873 8910 8925 882145 N/A N/A 9458 9473 CACGAAGGGCAGTGCC 83 1607 882168 N/A N/A 9747 9762 ATCTAATCCACAATCA 70 1608 882191 N/A N/A 10026 10041 CATTAAAGAGTGGTAC 103 1609 882215 N/A N/A 10199 10214 ACTAATTCCATAGAGC 21 1610 882239 N/A N/A 10519 10534 TGACATGCTTTGTCCT 41 1611 882263 N/A N/A 10795 10810 ATCAGATGATGTCTAG 103 1612 882287 N/A N/A 11160 11175 CTCCATTCAAGCCAGT 42 1613 882311 N/A N/A 11446 11461 GCAAGCTATATTAAAG 29 1614 882335 N/A N/A 11757 11772 CAAAAGTGCCACAACT 90 1615 882359 N/A N/A 12008 12023 ATCAGAAATCACGAGG 29 1616 882383 N/A N/A 12261 12276 ATCAATGGGAATCACA 10 1617 882406 N/A N/A 12701 12716 ACACACATTAAAGGGT 57 1618 882429 N/A N/A 13055 13070 AGCATCACTATTAAGA 28 1619 882453 N/A N/A 13323 13338 TGGAACTCTAGTGTCC 98 1620 882475 N/A N/A 14002 14017 ACCAGAATTGCAAGAG 40 1621 882497 N/A N/A 14216 14231 GTGCAAAACCAGTACA 67 1622 882520 N/A N/A 14640 14655 GAGCACATGTCAACAG 60 1623 882543 N/A N/A 15112 15127 TCACTTGCCGCCGCCT 51 1624 882567 N/A N/A 15600 15615 GTTCTAGTCGCGCAAG 73 1625 882590 N/A N/A 15855 15870 AATCAGAATGGCGAGT 61 1626 882613 N/A N/A 16052 16067 GTGCAGACCTGAATAG 86 1627 882637 N/A N/A 16269 16284 CTGGAACATTGTTGGA 54 1628 882660 N/A N/A 16501 16516 CAGAAAATATGTAACG 78 1629 882684 N/A N/A 16736 16751 CATTGGACCCACGAAG 72 1630 882708 N/A N/A 16975 16990 GTTCAGAAACCCTCAT 87 1631 882732 N/A N/A 17368 17383 GAAAGAGGCACCCTCC 81 1632 882756 N/A N/A 17597 17612 TAAAAGAGGTGCTAGG 73 1633 882780 N/A N/A 17889 17904 GTATGTAGCAATGTGA 79 1634 882804 N/A N/A 18126 18141 CTGGAGAGAACTCATG 62 1635 882827 N/A N/A 18687 18702 ATTTTATGGACCACGC 51 1636 882851 N/A N/A 19080 19095 CCACAACTGCAGTTTG 77 1637 882874 N/A N/A 19346 19361 AGCCCAAAGTTACATC 58 1638 882893 N/A N/A 4508 4523 CACTAAGTGGGCTCTG 59 1639

TABLE 24 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 23 195 881086 149 164 5176 5191 CCGAACTCTCCGCCTC 64 1640 881133 774 789 10839 10854 ATAAAAGGTTCCTGTC 16 1641 881180 1256 1271 17049 17064 TGGAATCTTGGCAGGG 60 1642 881204 1457 1472 19574 19589 ATAGAGGAATGGCGGA 35 1643 881228 1828 1843 19945 19960 AATCAGATGTCACTGA 97 1644 881252 1973 1988 20090 20105 CCTAATCTACAACAAA 85 1645 881276 2124 2139 20241 20256 CCTACATACATTAATA 107 1646 881300 2307 2322 20424 20439 TACTAACTGGCTTCCA 72 1647 881324 2402 2417 20519 20534 AACCATTACTTAGACA 55 1648 881348 2540 2555 20657 20672 CAGATAGTTAAGTACT 56 1649 881372 2760 2775 20877 20892 TGTCAAACTATAATTG 85 1650 881396 2840 2855 20957 20972 TAGGTATACTTAAGCA 21 1651 881420 2933 2948 21050 21065 GTAATAGTAAATATCC 39 1652 881444 3076 3091 21193 21208 CACTAAGCTTGATAAA 111 1653 881468 3377 3392 21494 21509 TGTTAGCTACAAAGAG 53 1654 881492 3456 3471 21573 21588 TGAAATGTCAAACCCT 46 1655 881515 3559 3574 21676 21691 GGATAATATACCCCAA 36 1656 881539 3731 3746 21848 21863 ATTGAAATGGAGCATC 101 1657 881587 4187 4202 22304 22319 AGGAGATATTAATTAC 43 1658 881606 4378 4393 22495 22510 GTAAGGGCTGCTCATT 48 1659 881630 4459 4474 22576 22591 GGCTAGCTGTATTTAT 63 1660 881654 4586 4601 22703 22718 TTACACTGGATCTGAA 62 1661 881678 4755 4770 22872 22887 TGTAAGGTCTGAGAGC 75 1662 881702 4978 4993 23095 23110 TCCATTGGAGGAGGGT 67 1663 881726 5184 5199 23301 23316 AGTTTGTGAAGCATCC 31 1664 881750 5222 5237 23339 23354 ATGATGTATACCAACG 14 1665 881774 N/A N/A 18807 18822 CATGATATTAGAGACT 44 1666 881798 N/A N/A 3865 3880 CTGGTTCGCGCTCCGG 93 1667 881819 N/A N/A 4244 4259 CCGGAGGCCGCCGTGA 87 1668 881842 N/A N/A 4510 4525 CGCACTAAGTGGGCTC 86 1669 881866 N/A N/A 4892 4907 GCACAGCCGTCCGCCT 84 1670 881890 N/A N/A 5492 5507 CGCCACCTGATGCCTC 70 1671 881913 N/A N/A 5726 5741 CCCACGGACGGAGGCG 109 1672 881934 N/A N/A 5896 5911 TGGGAGTACCCGCAGA 74 1673 881958 N/A N/A 6284 6299 ATAACCGGTAGAGTGA 50 1674 881980 N/A N/A 6683 6698 CCGCAGCATCGAACTC 58 1675 882028 N/A N/A 7481 7496 GGTCAGAATCTTGAAA 91 1676 882052 N/A N/A 7795 7810 AAACAGGAACTGACCT 80 1677 882076 N/A N/A 8220 8235 ATGATAAGGCACTACT 87 1678 882099 N/A N/A 8465 8480 AGCATTTTATCATCCG 19 1679 882123 N/A N/A 8781 8796 TTAAAGGAGTGCAGGA 84 1680 882146 N/A N/A 9460 9475 CCCACGAAGGGCAGTG 65 1681 882169 N/A N/A 9751 9766 CCACATCTAATCCACA 33 1682 882192 N/A N/A 10049 10064 GTGAACATGCCACTCA 38 1683 882216 N/A N/A 10200 10215 TACTAATTCCATAGAG 74 1684 882240 N/A N/A 10520 10535 ATGACATGCTTTGTCC 54 1685 882264 N/A N/A 10939 10954 AGCAAACCCTGCACTC 92 1686 882288 N/A N/A 11215 11230 GGTAAAGGCACATTCC 53 1687 882312 N/A N/A 11448 11463 TTGCAAGCTATATTAA 66 1688 882336 N/A N/A 11790 11805 TGGTAGGTCAAACTCC 113 1689 882360 N/A N/A 12009 12024 CATCAGAAATCACGAG 45 1690 882384 N/A N/A 12283 12298 GGTAACTGTATGGAAC 24 1691 882407 N/A N/A 12757 12772 GAATTAGGTGCTTAAT 41 1692 882430 N/A N/A 13060 13075 TATAGAGCATCACTAT 59 1693 882454 N/A N/A 13324 13339 GTGGAACTCTAGTGTC 88 1694 882476 N/A N/A 14021 14036 TAAGAGGCGACTGCTG 145 1695 882498 N/A N/A 14233 14248 CTCCTATAACTTCTCC 67 1696 882521 N/A N/A 14643 14658 TACGAGCACATGTCAA 64 1697 882544 N/A N/A 15146 15161 TAGAATGAGAGGTGTC 71 1698 882568 N/A N/A 15609 15624 TAATAGTAAGTTCTAG 100 1699 882591 N/A N/A 15860 15875 AGGCTAATCAGAATGG 58 1700 882614 N/A N/A 16099 16114 TCAGATACACACCCTC 63 1701 882638 N/A N/A 16276 16291 AAACGGACTGGAACAT 93 1702 882661 N/A N/A 16516 16531 GTAACTCAGGCACCAC 53 1703 882685 N/A N/A 16758 16773 GCTAAAGAACACTGCT 97 1704 882709 N/A N/A 16981 16996 GACCATGTTCAGAAAC 4 1705 882733 N/A N/A 17369 17384 GGAAAGAGGCACCCTC 72 1706 882757 N/A N/A 17640 17655 TATCATATGCCCAATA 68 1707 882781 N/A N/A 17894 17909 TGCAAGTATGTAGCAA 97 1708 882805 N/A N/A 18131 18146 AATCACTGGAGAGAAC 71 1709 882828 N/A N/A 18688 18703 CATTTTATGGACCACG 38 1710 882852 N/A N/A 19099 19114 GGCCAATGATTTTGTT 94 1711 882875 N/A N/A 19350 19365 GTAAAGCCCAAAGTTA 73 1712

Example 6: Effect of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured SK-MEL-28 cells at a density of 20,000 cells per well were transfected using electroporation with 4,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 25 through 36 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 25 through 36 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 25 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 44 195 881087 151 166 5178 5193 TGCCGAACTCTCCGCC 57 1713 881110 482 497 6961 6976 ACTTTGTACGGGTCTG 61 1714 881134 778 793 10843 10858 AAGCATAAAAGGTTCC 72 1715 881157 1059 1074 13612 13627 GACCACGCCCCTCTCC 45 1716 881181 1263 1278 17056 17071 AGTCACCTGGAATCTT 53 1717 881205 1458 1473 19575 19590 AATAGAGGAATGGCGG 47 1718 881229 1831 1846 19948 19963 GCCAATCAGATGTCAC 54 1719 881253 1974 1989 20091 20106 ACCTAATCTACAACAA 83 1720 881277 2128 2143 20245 20260 AGCTCCTACATACATT 56 1721 881301 2309 2324 20426 20441 TTTACTAACTGGCTTC 55 1722 881325 2432 2447 20549 20564 GGTCAAAAAGATGCAG 49 1723 881349 2541 2556 20658 20673 ACAGATAGTTAAGTAC 65 1724 881373 2770 2785 20887 20902 TTTAAGGCCCTGTCAA 117 1725 881397 2842 2857 20959 20974 GATAGGTATACTTAAG 49 1726 881421 2938 2953 21055 21070 TGGGAGTAATAGTAAA 84 1727 881445 3077 3092 21194 21209 TCACTAAGCTTGATAA 63 1728 881469 3383 3398 21500 21515 CTTCACTGTTAGCTAC 49 1729 881493 3468 3483 21585 21600 TTGCATGGCTAATGAA 58 1730 881516 3560 3575 21677 21692 AGGATAATATACCCCA 26 1731 881540 3733 3748 21850 21865 CAATTGAAATGGAGCA 74 1732 881564 3956 3971 22073 22088 CCTGAGACAAGTGCTA 57 1733 881588 4200 4215 22317 22332 TCTATAGTGTTCCAGG 20 1734 881607 4380 4395 22497 22512 CTGTAAGGGCTGCTCA 44 1735 881631 4482 4497 22599 22614 TCCCAGAGTTGTTCCA 74 1736 881655 4587 4602 22704 22719 TTTACACTGGATCTGA 75 1737 881679 4756 4771 22873 22888 GTGTAAGGTCTGAGAG 84 1738 881703 4979 4994 23096 23111 TTCCATTGGAGGAGGG 72 1739 881727 5185 5200 23302 23317 CAGTTTGTGAAGCATC 22 1740 881751 5223 5238 23340 23355 CATGATGTATACCAAC 49 1741 881775 N/A N/A 18808 18823 TCATGATATTAGAGAC 65 1742 881799 N/A N/A 3874 3889 GTCGAACCTCTGGTTC 110 1743 881820 N/A N/A 4246 4261 CGCCGGAGGCCGCCGT 97 1744 881843 N/A N/A 4511 4526 GCGCACTAAGTGGGCT 110 1745 881867 N/A N/A 4893 4908 GGCACAGCCGTCCGCC 89 1746 881891 N/A N/A 5535 5550 CTGAGAGCCGAGGCCT 117 1747 881914 N/A N/A 5727 5742 ACCCACGGACGGAGGC 109 1748 881935 N/A N/A 5903 5918 ACAGAGGTGGGAGTAC 174 1749 881959 N/A N/A 6285 6300 TATAACCGGTAGAGTG 79 1750 881981 N/A N/A 6697 6712 ACGATGATAGCTCACC 92 1751 882005 N/A N/A 7221 7236 TACATTGAGGCATACA 59 1752 882029 N/A N/A 7547 7562 CCCAAGTGAGGTCACC 80 1753 882053 N/A N/A 7837 7852 GGCTCCTACATGTTTG 146 1754 882077 N/A N/A 8222 8237 ACATGATAAGGCACTA 39 1755 882100 N/A N/A 8470 8485 GCCGAAGCATTTTATC 71 1756 882124 N/A N/A 8782 8797 GTTAAAGGAGTGCAGG 114 1757 882147 N/A N/A 9461 9476 TCCCACGAAGGGCAGT 94 1758 882170 N/A N/A 9788 9803 TCATTTTGATGTCTGG 37 1759 882193 N/A N/A 10078 10093 GCCAATGCAACTGAAT 64 1760 882217 N/A N/A 10202 10217 GTTACTAATTCCATAG 51 1761 882241 N/A N/A 10525 10540 GACAGATGACATGCTT 91 1762 882265 N/A N/A 10940 10955 GAGCAAACCCTGCACT 116 1763 882289 N/A N/A 11216 11231 AGGTAAAGGCACATTC 81 1764 882313 N/A N/A 11469 11484 TCAGATTGAATCCATA 36 1765 882337 N/A N/A 11793 11808 AGCTGGTAGGTCAAAC 105 1766 882361 N/A N/A 12050 12065 TTCGAGGTGATTCTCG 95 1767 882385 N/A N/A 12284 12299 TGGTAACTGTATGGAA 52 1768 882408 N/A N/A 12758 12773 AGAATTAGGTGCTTAA 39 1769 882431 N/A N/A 13064 13079 CTATTATAGAGCATCA 45 1770 882455 N/A N/A 13356 13371 GGCCAACGACTCCACA 77 1771 882477 N/A N/A 14022 14037 TTAAGAGGCGACTGCT 88 1772 882499 N/A N/A 14244 14259 CTTATAGCACTCTCCT 67 1773 882522 N/A N/A 14645 14660 AGTACGAGCACATGTC 66 1774 882545 N/A N/A 15191 15206 AAGGATGGGACCGCCC 61 1775 882569 N/A N/A 15613 15628 AGATTAATAGTAAGTT 96 1776 882592 N/A N/A 15864 15879 ACACAGGCTAATCAGA 93 1777 882615 N/A N/A 16100 16115 ATCAGATACACACCCT 108 1778 882639 N/A N/A 16278 16293 CAAAACGGACTGGAAC 84 1779 882662 N/A N/A 16517 16532 CGTAACTCAGGCACCA 70 1780 882686 N/A N/A 16762 16777 GACAGCTAAAGAACAC 81 1781 882710 N/A N/A 16993 17008 CCACAAGAAAGAGACC 109 1782 882734 N/A N/A 17400 17415 CGACAACTTTCCTGAA 79 1783 882758 N/A N/A 17644 17659 CAAATATCATATGCCC 31 1784 882782 N/A N/A 17904 17919 TAATAATGCTTGCAAG 88 1785 882806 N/A N/A 18135 18150 AGTCAATCACTGGAGA 40 1786 882829 N/A N/A 18691 18706 ATTCATTTTATGGACC 58 1787 882853 N/A N/A 19108 19123 GGTAATTTAGGCCAAT 65 1788 882876 N/A N/A 19366 19381 CTAAGGAGACAGTAAC 71 1789

TABLE 26 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 42 195 881088 157 172 5184 5199 CGCTCATGCCGAACTC 89 1790 881111 485 500 6964 6979 TACACTTTGTACGGGT 72 1791 881135 782 797 10847 10862 GCACAAGCATAAAAGG 84 1792 881158 1062 1077 13615 13630 GAGGACCACGCCCCTC 103 1793 881182 1269 1284 17062 17077 GCATAGAGTCACCTGG 61 1794 881206 1459 1474 19576 19591 GAATAGAGGAATGGCG 61 1795 881230 1832 1847 19949 19964 TGCCAATCAGATGTCA 78 1796 881254 1975 1990 20092 20107 GACCTAATCTACAACA 67 1797 881278 2150 2165 20267 20282 AAGTGTCTTCCACAAG 46 1798 881302 2310 2325 20427 20442 GTTTACTAACTGGCTT 73 1799 881326 2443 2458 20560 20575 TAAAGAATGAGGGTCA 55 1800 881350 2543 2558 20660 20675 GAACAGATAGTTAAGT 83 1801 881374 2771 2786 20888 20903 TTTTAAGGCCCTGTCA 82 1802 881398 2843 2858 20960 20975 TGATAGGTATACTTAA 72 1803 881422 2957 2972 21074 21089 CGCAATCTTCTGCTGA 36 1804 881446 3079 3094 21196 21211 GCTCACTAAGCTTGAT 44 1805 881470 3390 3405 21507 21522 GGTAAATCTTCACTGT 35 1806 881494 3475 3490 21592 21607 ATCCATGTTGCATGGC 39 1807 881517 3561 3576 21678 21693 TAGGATAATATACCCC 26 1808 881541 3734 3749 21851 21866 GCAATTGAAATGGAGC 76 1809 881565 3979 3994 22096 22111 AGGAAGCCGTTCCTTT 58 1810 881589 4201 4216 22318 22333 CTCTATAGTGTTCCAG 34 1811 881608 4381 4396 22498 22513 ACTGTAAGGGCTGCTC 31 1812 881632 4495 4510 22612 22627 GAGTACCCAAGACTCC 75 1813 881656 4588 4603 22705 22720 GTTTACACTGGATCTG 39 1814 881680 4765 4780 22882 22897 CAAAATGGTGTGTAAG 75 1815 881704 4983 4998 23100 23115 GAATTTCCATTGGAGG 57 1816 881728 5187 5202 23304 23319 CTCAGTTTGTGAAGCA 15 1817 881752 5224 5239 23341 23356 TCATGATGTATACCAA 35 1818 881776 N/A N/A 18810 18825 AATCATGATATTAGAG 69 1819 881800 N/A N/A 3880 3895 CTGGAGGTCGAACCTC 98 1820 881821 N/A N/A 4257 4272 GGTGAGCACCGCGCCG 108 1821 881844 N/A N/A 4522 4537 GCCCAGCTAGCGCGCA 71 1822 881868 N/A N/A 4909 4924 GCGCATCCCCTGGGCG 104 1823 881892 N/A N/A 5538 5553 CCGCTGAGAGCCGAGG 104 1824 881915 N/A N/A 5730 5745 GGGACCCACGGACGGA 91 1825 881936 N/A N/A 5972 5987 CTCTAGACAGAGGCCC 80 1826 881960 N/A N/A 6286 6301 TTATAACCGGTAGAGT 62 1827 881982 N/A N/A 6761 6776 GGTTTATTAGCTTTCT 101 1828 882006 N/A N/A 7226 7241 CCATATACATTGAGGC 56 1829 882030 N/A N/A 7574 7589 ATAAAGGACCCCGCCA 82 1830 882054 N/A N/A 8019 8034 GGCAAGTTCTGCTGTC 42 1831 882078 N/A N/A 8226 8241 TTTCACATGATAAGGC 57 1832 882101 N/A N/A 8471 8486 AGCCGAAGCATTTTAT 56 1833 882125 N/A N/A 9093 9108 CTAAAGGAGTGCAGGA 94 1834 882148 N/A N/A 9467 9482 ATAAGATCCCACGAAG 142 1835 882171 N/A N/A 9816 9831 AGCTAATGAGAGCTTC 83 1836 882194 N/A N/A 10086 10101 CTTGAAAAGCCAATGC 42 1837 882218 N/A N/A 10203 10218 GGTTACTAATTCCATA 44 1838 882242 N/A N/A 10526 10541 AGACAGATGACATGCT 50 1839 882266 N/A N/A 11013 11028 TCTAAAGTCCCATCGA 69 1840 882290 N/A N/A 11220 11235 GTAAAGGTAAAGGCAC 58 1841 882314 N/A N/A 11528 11543 GGTAAGATCTCCATGG 55 1842 882338 N/A N/A 11798 11813 GAAAGAGCTGGTAGGT 100 1843 882362 N/A N/A 12054 12069 GCCATTCGAGGTGATT 44 1844 882386 N/A N/A 12355 12370 ACCAAGCTGGGTTTGC 47 1845 882409 N/A N/A 12760 12775 ATAGAATTAGGTGCTT 36 1846 882432 N/A N/A 13065 13080 CCTATTATAGAGCATC 32 1847 882456 N/A N/A 13361 13376 CTCGAGGCCAACGACT 99 1848 882478 N/A N/A 14025 14040 GTTTTAAGAGGCGACT 67 1849 882500 N/A N/A 14246 14261 AACTTATAGCACTCTC 72 1850 882523 N/A N/A 14648 14663 AGAAGTACGAGCACAT 56 1851 882546 N/A N/A 15192 15207 GAAGGATGGGACCGCC 59 1852 882570 N/A N/A 15617 15632 TCACAGATTAATAGTA 77 1853 882593 N/A N/A 15867 15882 CCTACACAGGCTAATC 87 1854 882616 N/A N/A 16101 16116 TATCAGATACACACCC 81 1855 882640 N/A N/A 16279 16294 ACAAAACGGACTGGAA 86 1856 882663 N/A N/A 16535 16550 TGCTACTGCGGACATC 57 1857 882687 N/A N/A 16763 16778 CGACAGCTAAAGAACA 63 1858 882711 N/A N/A 17000 17015 TAGAAGCCCACAAGAA 99 1859 882735 N/A N/A 17402 17417 TACGACAACTTTCCTG 59 1860 882759 N/A N/A 17662 17677 GAAAGATTCAGCCTCT 55 1861 882783 N/A N/A 17906 17921 GTTAATAATGCTTGCA 98 1862 882807 N/A N/A 18141 18156 TTATTAAGTCAATCAC 101 1863 882830 N/A N/A 18714 18729 CTCATAGGTGTACACG 41 1864 882854 N/A N/A 19114 19129 CTGCAGGGTAATTTAG 66 1865 882877 N/A N/A 19379 19394 CAGCACTCAGATTCTA 88 1866

TABLE 27 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 41 195 668855 783 798 10848 10863 GGCACAAGCATAAAAG 99 1867 690520 4222 4237 22339 22354 AAATGAGTCGGTCACT 45 1868 881089 165 180 5192 5207 GCTCACCGCGCTCATG 52 1869 881112 486 501 6965 6980 GTACACTTTGTACGGG 36 1870 881159 1067 1082 13620 13635 ATCCAGAGGACCACGC 81 1871 881183 1270 1285 17063 17078 AGCATAGAGTCACCTG 37 1872 881207 1460 1475 19577 19592 TGAATAGAGGAATGGC 79 1873 881231 1844 1859 19961 19976 AATAAGCTCATCTGCC 64 1874 881255 1978 1993 20095 20110 CAAGACCTAATCTACA 88 1875 881279 2151 2166 20268 20283 CAAGTGTCTTCCACAA 35 1876 881303 2311 2326 20428 20443 AGTTTACTAACTGGCT 37 1877 881327 2444 2459 20561 20576 CTAAAGAATGAGGGTC 38 1878 881351 2545 2560 20662 20677 GGGAACAGATAGTTAA 74 1879 881375 2773 2788 20890 20905 AATTTTAAGGCCCTGT 76 1880 881399 2844 2859 20961 20976 GTGATAGGTATACTTA 35 1881 881423 2958 2973 21075 21090 ACGCAATCTTCTGCTG 48 1882 881447 3086 3101 21203 21218 GCTCACTGCTCACTAA 67 1883 881471 3391 3406 21508 21523 AGGTAAATCTTCACTG 46 1884 881495 3479 3494 21596 21611 ACATATCCATGTTGCA 21 1885 881518 3562 3577 21679 21694 TTAGGATAATATACCC 48 1886 881542 3785 3800 21902 21917 ACTGTTAAAGCAGCAT 29 1887 881566 3980 3995 22097 22112 GAGGAAGCCGTTCCTT 49 1888 881609 4383 4398 22500 22515 ATACTGTAAGGGCTGC 46 1889 881633 4504 4519 22621 22636 AAGAGGTGCGAGTACC 66 1890 881657 4590 4605 22707 22722 AAGTTTACACTGGATC 42 1891 881681 4786 4801 22903 22918 GGGCATGTAAAACATA 77 1892 881705 4985 5000 23102 23117 GGGAATTTCCATTGGA 56 1893 881729 5188 5203 23305 23320 CCTCAGTTTGTGAAGC 31 1894 881753 5226 5241 23343 23358 ATTCATGATGTATACC 38 1895 881777 N/A N/A 18811 18826 CAATCATGATATTAGA 62 1896 881801 N/A N/A 3897 3912 GGGTACCCTGCGCTGC 56 1897 881822 N/A N/A 4292 4307 CCAAATGTGGAGCTCC 77 1898 881845 N/A N/A 4542 4557 CGAATAGGACCCCTAT 91 1899 881869 N/A N/A 4916 4931 GGCCCGGGCGCATCCC 95 1900 881893 N/A N/A 5548 5563 CCCCGCGGTCCCGCTG 59 1901 881916 N/A N/A 5749 5764 ACGCACGGAGAGGGCG 99 1902 881937 N/A N/A 5974 5989 AGCTCTAGACAGAGGC 65 1903 881961 N/A N/A 6287 6302 TTTATAACCGGTAGAG 46 1904 881983 N/A N/A 6783 6798 CGAAAAGTCAAAATAC 100 1905 882007 N/A N/A 7228 7243 CCCCATATACATTGAG 63 1906 882031 N/A N/A 7576 7591 ACATAAAGGACCCCGC 72 1907 882055 N/A N/A 8027 8042 TAGCAAATGGCAAGTT 83 1908 882079 N/A N/A 8248 8263 GAAGAGATCAGCTGCC 58 1909 882102 N/A N/A 8475 8490 TGACAGCCGAAGCATT 62 1910 882126 N/A N/A 9094 9109 GCTAAAGGAGTGCAGG 93 1911 882149 N/A N/A 9468 9483 AATAAGATCCCACGAA 85 1912 882172 N/A N/A 9820 9835 ACCCAGCTAATGAGAG 65 1913 882195 N/A N/A 10098 10113 CTGCAAATCCCTCTTG 133 1914 882219 N/A N/A 10230 10245 CAGTTCTAAGCATTGC 67 1915 882243 N/A N/A 10551 10566 GACTAACAGGGAGACT 109 1916 882267 N/A N/A 11014 11029 GTCTAAAGTCCCATCG 60 1917 882291 N/A N/A 11251 11266 CGTGAGAATGTTGGCT 51 1918 882315 N/A N/A 11533 11548 TAGTAGGTAAGATCTC 77 1919 882339 N/A N/A 11799 11814 AGAAAGAGCTGGTAGG 64 1920 882363 N/A N/A 12074 12089 CCCAAAGAGAGTGGGT 100 1921 882387 N/A N/A 12364 12379 GTTGAAAGAACCAAGC 89 1922 882410 N/A N/A 12761 12776 TATAGAATTAGGTGCT 60 1923 882433 N/A N/A 13112 13127 GGAACAAGTGTATCTT 24 1924 882457 N/A N/A 13367 13382 CACCACCTCGAGGCCA 76 1925 882479 N/A N/A 14027 14042 CTGTTTTAAGAGGCGA 42 1926 882501 N/A N/A 14250 14265 AGCCAACTTATAGCAC 46 1927 882524 N/A N/A 14649 14664 CAGAAGTACGAGCACA 53 1928 882547 N/A N/A 15195 15210 CAAGAAGGATGGGACC 75 1929 882571 N/A N/A 15619 15634 GCTCACAGATTAATAG 84 1930 882594 N/A N/A 15871 15886 TACACCTACACAGGCT 81 1931 882617 N/A N/A 16121 16136 CTGCAGAACAGACGCG 89 1932 882641 N/A N/A 16280 16295 TACAAAACGGACTGGA 62 1933 882664 N/A N/A 16551 16566 CATAATCCAGTATCTG 82 1934 882688 N/A N/A 16767 16782 TAGTCGACAGCTAAAG 54 1935 882712 N/A N/A 17001 17016 GTAGAAGCCCACAAGA 104 1936 882736 N/A N/A 17404 17419 AATACGACAACTTTCC 50 1937 882760 N/A N/A 17664 17679 TTGAAAGATTCAGCCT 97 1938 882784 N/A N/A 17918 17933 GCATGCAAGCCCGTTA 101 1939 882808 N/A N/A 18183 18198 TTGTTAACAATGTATC 53 1940 882831 N/A N/A 18715 18730 ACTCATAGGTGTACAC 55 1941 882855 N/A N/A 19126 19141 GTCTGAGGGAATCTGC 51 1942 882878 N/A N/A 19384 19399 TTAAACAGCACTCAGA 67 1943

TABLE 28 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 20 195 690521 4224 4239 22341 22356 GTAAATGAGTCGGTCA 21 1944 881090 170 185 5197 5212 CCGCAGCTCACCGCGC 64 1945 881113 488 503 6967 6982 CTGTACACTTTGTACG 66 1946 881136 837 852 10902 10917 GGCAGACCTTATGCTT 79 1947 881160 1071 1086 13624 13639 GGCCATCCAGAGGACC 75 1948 881184 1271 1286 17064 17079 AAGCATAGAGTCACCT 64 1949 881208 1461 1476 19578 19593 TTGAATAGAGGAATGG 58 1950 881232 1886 1901 20003 20018 GTCTACAGAACACAAG 76 1951 881256 1984 1999 20101 20116 TTCCAGCAAGACCTAA 73 1952 881280 2152 2167 20269 20284 GCAAGTGTCTTCCACA 23 1953 881304 2312 2327 20429 20444 AAGTTTACTAACTGGC 37 1954 881328 2462 2477 20579 20594 CGAAGAATTTTAGCAT 42 1955 881352 2546 2561 20663 20678 AGGGAACAGATAGTTA 66 1956 881376 2774 2789 20891 20906 TAATTTTAAGGCCCTG 61 1957 881400 2845 2860 20962 20977 AGTGATAGGTATACTT 61 1958 881424 2962 2977 21079 21094 AGCTACGCAATCTTCT 53 1959 881448 3250 3265 21367 21382 ACTTTTAGAGAGGAGA 36 1960 881472 3394 3409 21511 21526 ACGAGGTAAATCTTCA 39 1961 881496 3480 3495 21597 21612 TACATATCCATGTTGC 26 1962 881519 3564 3579 21681 21696 CCTTAGGATAATATAC 77 1963 881543 3787 3802 21904 21919 CCACTGTTAAAGCAGC 34 1964 881567 3983 3998 22100 22115 AATGAGGAAGCCGTTC 53 1965 881610 4384 4399 22501 22516 AATACTGTAAGGGCTG 36 1966 881634 4506 4521 22623 22638 CCAAGAGGTGCGAGTA 67 1967 881658 4591 4606 22708 22723 GAAGTTTACACTGGAT 35 1968 881682 4802 4817 22919 22934 ATCAGTCTCAAAAACG 59 1969 881706 4989 5004 23106 23121 ACACGGGAATTTCCAT 44 1970 881730 5189 5204 23306 23321 ACCTCAGTTTGTGAAG 61 1971 881754 5248 5263 23365 23380 TTGCAGAGCAATTTAC 50 1972 881778 N/A N/A 18814 18829 CATCAATCATGATATT 75 1973 881823 N/A N/A 4304 4319 GCTCGGAGCGACCCAA 92 1974 881846 N/A N/A 4543 4558 CCGAATAGGACCCCTA 68 1975 881870 N/A N/A 4920 4935 GGCCGGCCCGGGCGCA 99 1976 881894 N/A N/A 5566 5581 CAGGACCCGGCTCCCG 86 1977 881917 N/A N/A 5752 5767 CGGACGCACGGAGAGG 72 1978 881938 N/A N/A 5982 5997 AGGGAGACAGCTCTAG 75 1979 881962 N/A N/A 6290 6305 GTATTTATAACCGGTA 33 1980 881984 N/A N/A 7014 7029 CAAATTCAGGAGAGCC 81 1981 882008 N/A N/A 7247 7262 ACATATTCAATGCACC 53 1982 882032 N/A N/A 7578 7593 TGACATAAAGGACCCC 52 1983 882056 N/A N/A 8032 8047 AGCCATAGCAAATGGC 95 1984 882080 N/A N/A 8302 8317 CTTTTACCCACCAAAG 82 1985 882103 N/A N/A 8481 8496 TTAGACTGACAGCCGA 47 1986 882127 N/A N/A 9278 9293 CAATTAGCTCTTCTAT 86 1987 882150 N/A N/A 9471 9486 TTAAATAAGATCCCAC 77 1988 882173 N/A N/A 9835 9850 CTAGATTCTCCCTGCA 66 1989 882196 N/A N/A 10110 10125 AACCAGCCCTTGCTGC 79 1990 882220 N/A N/A 10237 10252 CTTTACACAGTTCTAA 113 1991 882244 N/A N/A 10554 10569 ACTGACTAACAGGGAG 51 1992 882268 N/A N/A 11021 11036 AGCAAGTGTCTAAAGT 38 1993 882292 N/A N/A 11271 11286 TCCAAGCAATTATTCC 88 1994 882316 N/A N/A 11534 11549 ATAGTAGGTAAGATCT 113 1995 882340 N/A N/A 11800 11815 TAGAAAGAGCTGGTAG 91 1996 882364 N/A N/A 12075 12090 CCCCAAAGAGAGTGGG 111 1997 882388 N/A N/A 12372 12387 GCCCATGAGTTGAAAG 68 1998 882411 N/A N/A 12762 12777 ATATAGAATTAGGTGC 84 1999 882434 N/A N/A 13113 13128 AGGAACAAGTGTATCT 54 2000 882502 N/A N/A 14254 14269 ACAGAGCCAACTTATA 94 2001 882525 N/A N/A 14652 14667 ACCCAGAAGTACGAGC 79 2002 882548 N/A N/A 15218 15233 CCCATGAACACCATGC 68 2003 882572 N/A N/A 15635 15650 GCCAAGACCAGCTCTT 70 2004 882595 N/A N/A 15872 15887 CTACACCTACACAGGC 84 2005 882618 N/A N/A 16132 16147 CGCTACAGCTTCTGCA 80 2006 882642 N/A N/A 16281 16296 ATACAAAACGGACTGG 85 2007 882665 N/A N/A 16553 16568 CACATAATCCAGTATC 57 2008 882689 N/A N/A 16773 16788 AGGAACTAGTCGACAG 60 2009 882713 N/A N/A 17133 17148 TGTAACTGAGGACTCA 102 2010 882737 N/A N/A 17405 17420 AAATACGACAACTTTC 79 2011 882761 N/A N/A 17676 17691 TGCTTGAATTCCTTGA 33 2012 882785 N/A N/A 17923 17938 GCTCAGCATGCAAGCC 100 2013 882809 N/A N/A 18225 18240 GCTTATCAATGCCAAG 60 2014 882832 N/A N/A 18716 18731 AACTCATAGGTGTACA 39 2015 882856 N/A N/A 19154 19169 TCAAGTAAAGTGATCA 52 2016 882879 N/A N/A 19387 19402 CTATTAAACAGCACTC 82 2017 882890 N/A N/A 3949 3964 GTGGGAGTCGGAGCTC 83 2018 882906 N/A N/A 13375 13390 GCCAAGGACACCACCT 102 2019 882908 N/A N/A 14061 14076 GTATTTGTCGAGATCA 41 2020

TABLE 29 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: Compound 1 Start 1 Stop 2 Start 2 Stop IRF4 (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 33 195 690522 4228 4243 22345 22360 AGTTGTAAATGAGTCG 15 2021 881091 186 201 5213 5228 GCGGAGCTTCCCGTTG 67 2022 881114 495 510 6974 6989 AACAATCCTGTACACT 78 2023 881161 1088 1103 13641 13656 GCATAGAGCCCGTCGG 50 2024 881185 1272 1287 17065 17080 AAAGCATAGAGTCACC 66 2025 881209 1532 1547 19649 19664 CCGTATCCCCGTATCA 65 2026 881233 1891 1906 20008 20023 TGGCAGTCTACAGAAC 122 2027 881257 2001 2016 20118 20133 GGCAAGTTTTCTCTGT 64 2028 881281 2154 2169 20271 20286 CAGCAAGTGTCTTCCA 36 2029 881305 2313 2328 20430 20445 GAAGTTTACTAACTGG 43 2030 881329 2463 2478 20580 20595 GCGAAGAATTTTAGCA 88 2031 881353 2547 2562 20664 20679 AAGGGAACAGATAGTT 80 2032 881377 2775 2790 20892 20907 GTAATTTTAAGGCCCT 45 2033 881401 2846 2861 20963 20978 AAGTGATAGGTATACT 50 2034 881425 2965 2980 21082 21097 GAGAGCTACGCAATCT 43 2035 881449 3251 3266 21368 21383 CACTTTTAGAGAGGAG 38 2036 881473 3395 3410 21512 21527 AACGAGGTAAATCTTC 49 2037 881497 3484 3499 21601 21616 CCAATACATATCCATG 40 2038 881520 3566 3581 21683 21698 TCCCTTAGGATAATAT 52 2039 881544 3788 3803 21905 21920 TCCACTGTTAAAGCAG 45 2040 881568 3984 3999 22101 22116 GAATGAGGAAGCCGTT 61 2041 881611 4385 4400 22502 22517 CAATACTGTAAGGGCT 44 2042 881635 4507 4522 22624 22639 GCCAAGAGGTGCGAGT 65 2043 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA 15 2044 881683 4808 4823 22925 22940 ATCGAGATCAGTCTCA 24 2045 881707 4991 5006 23108 23123 CAACACGGGAATTTCC 45 2046 881731 5191 5206 23308 23323 CTACCTCAGTTTGTGA 59 2047 881755 N/A N/A 18767 18782 CTAAACTCCTAAGTAC 86 2048 881779 N/A N/A 18827 18842 GCTGAATGCTCTCCAT 48 2049 881802 N/A N/A 3965 3980 GCTCAGCGCAGATGGG 102 2050 881824 N/A N/A 4310 4325 CGCAAGGCTCGGAGCG 90 2051 881847 N/A N/A 4544 4559 CCCGAATAGGACCCCT 60 2052 881871 N/A N/A 4932 4947 AGGCACCTTCGCGGCC 82 2053 881918 N/A N/A 5754 5769 CGCGGACGCACGGAGA 81 2054 881939 N/A N/A 5995 6010 CGCGCAAAGGGCAAGG 66 2055 881963 N/A N/A 6292 6307 GGGTATTTATAACCGG 41 2056 881985 N/A N/A 7032 7047 TGCCTCTGTTAGGTGA 66 2057 882009 N/A N/A 7268 7283 TTGAAAGACTAACTGG 80 2058 882033 N/A N/A 7580 7595 TGTGACATAAAGGACC 73 2059 882057 N/A N/A 8040 8055 GTTGGAGCAGCCATAG 69 2060 882081 N/A N/A 8329 8344 CAAAAGTACCACAGGG 60 2061 882104 N/A N/A 8484 8499 TTATTAGACTGACAGC 63 2062 882128 N/A N/A 9279 9294 GCAATTAGCTCTTCTA 79 2063 882151 N/A N/A 9525 9540 CTATATAAAAAGTGGG 84 2064 882174 N/A N/A 9837 9852 TGCTAGATTCTCCCTG 67 2065 882197 N/A N/A 10114 10129 TGGAAACCAGCCCTTG 52 2066 882221 N/A N/A 10250 10265 ACCTACTCCATCACTT 103 2067 882245 N/A N/A 10559 10574 CTGTAACTGACTAACA 77 2068 882269 N/A N/A 11022 11037 AAGCAAGTGTCTAAAG 71 2069 882293 N/A N/A 11278 11293 CTACACTTCCAAGCAA 105 2070 882317 N/A N/A 11535 11550 TATAGTAGGTAAGATC 87 2071 882341 N/A N/A 11801 11816 CTAGAAAGAGCTGGTA 85 2072 882365 N/A N/A 12087 12102 TCAGACAGTGCGCCCC 74 2073 882389 N/A N/A 12379 12394 AAATATCGCCCATGAG 61 2074 882412 N/A N/A 12763 12778 TATATAGAATTAGGTG 116 2075 882435 N/A N/A 13115 13130 TAAGGAACAAGTGTAT 82 2076 882458 N/A N/A 13393 13408 CCGGAGTCAGTGCTGG 107 2077 882480 N/A N/A 14062 14077 CGTATTTGTCGAGATC 50 2078 882503 N/A N/A 14256 14271 TCACAGAGCCAACTTA 112 2079 882526 N/A N/A 14656 14671 TTACACCCAGAAGTAC 95 2080 882549 N/A N/A 15312 15327 GCCCATGTGAGCTCTT 50 2081 882573 N/A N/A 15704 15719 GAACACTTTGAGGTGA 86 2082 882596 N/A N/A 15874 15889 GACTACACCTACACAG 68 2083 882619 N/A N/A 16135 16150 AGCCGCTACAGCTTCT 108 2084 882643 N/A N/A 16282 16297 GATACAAAACGGACTG 56 2085 882666 N/A N/A 16556 16571 GCCCACATAATCCAGT 78 2086 882690 N/A N/A 16774 16789 GAGGAACTAGTCGACA 61 2087 882714 N/A N/A 17139 17154 TAGGAGTGTAACTGAG 63 2088 882738 N/A N/A 17406 17421 GAAATACGACAACTTT 78 2089 882762 N/A N/A 17693 17708 TACGAGAGGGTCTGAT 100 2090 882786 N/A N/A 17936 17951 TTTACCTGGTATTGCT 62 2091 882810 N/A N/A 18226 18241 TGCTTATCAATGCCAA 30 2092 882833 N/A N/A 18717 18732 GAACTCATAGGTGTAC 33 2093 882880 N/A N/A 19389 19404 CACTATTAAACAGCAC 81 2094 882916 N/A N/A 19155 19170 ATCAAGTAAAGTGATC 71 2095

TABLE 30 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  29  195 690523 4230 4245 22347 22362 TCAGTTGTAAATGAGT  28 2096 881092  187  202  5214  5229 GGCGGAGCTTCCCGTT  74 2097 881115  498  513  6977  6992 AGGAACAATCCTGTAC  84 2098 881162 1089 1104 13642 13657 CGCATAGAGCCCGTCG  47 2099 881186 1273 1288 17066 17081 CAAAGCATAGAGTCAC  64 2100 881210 1534 1549 19651 19666 CCCCGTATCCCCGTAT  55 2101 881234 1896 1911 20013 20028 AATGATGGCAGTCTAC  67 2102 881258 2014 2029 20131 20146 GTCAATACTGAAAGGC  34 2103 881282 2155 2170 20272 20287 TCAGCAAGTGTCTTCC  34 2104 881306 2316 2331 20433 20448 TAGGAAGTTTACTAAC  64 2105 881330 2464 2479 20581 20596 TGCGAAGAATTTTAGC  59 2106 881354 2548 2563 20665 20680 GAAGGGAACAGATAGT  83 2107 881378 2776 2791 20893 20908 AGTAATTTTAAGGCCC  40 2108 881402 2847 2862 20964 20979 TAAGTGATAGGTATAC  40 2109 881426 2972 2987 21089 21104 CACATTTGAGAGCTAC  26 2110 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA  37 2111 881474 3396 3411 21513 21528 GAACGAGGTAAATCTT  44 2112 881498 3485 3500 21602 21617 CCCAATACATATCCAT  26 2113 881521 3600 3615 21717 21732 CGTGAAACAGCAGTTC  60 2114 881545 3789 3804 21906 21921 CTCCACTGTTAAAGCA  48 2115 881569 3986 4001 22103 22118 AGGAATGAGGAAGCCG  44 2116 881612 4387 4402 22504 22519 AACAATACTGTAAGGG  49 2117 881636 4508 4523 22625 22640 AGCCAAGAGGTGCGAG  62 2118 881660 4593 4608 22710 22725 CGGAAGTTTACACTGG  15 2119 881684 4809 4824 22926 22941 CATCGAGATCAGTCTC  35 2120 881708 4993 5008 23110 23125 AGCAACACGGGAATTT  43 2121 881732 5197 5212 23314 23329 CATTATCTACCTCAGT  65 2122 881756 N/A N/A 18768 18783 GCTAAACTCCTAAGTA  70 2123 881780 N/A N/A 18828 18843 AGCTGAATGCTCTCCA  22 2124 881803 N/A N/A  3993  4008 GGCCTTGGAGCCCAAA  80 2125 881825 N/A N/A  4311  4326 ACGCAAGGCTCGGAGC  79 2126 881848 N/A N/A  4545  4560 CCCCGAATAGGACCCC  44 2127 881872 N/A N/A  4937  4952 GAAGAAGGCACCTTCG  75 2128 881896 N/A N/A  5646  5661 GCAAAACCCCTCAAGC  80 2129 881919 N/A N/A  5755  5770 GCGCGGACGCACGGAG  98 2130 881940 N/A N/A  6001  6016 TGCACTCGCGCAAAGG  72 2131 881964 N/A N/A  6315  6330 GCCAAGTTGAAGACAC  43 2132 881986 N/A N/A  7057  7072 AGTTAAGGTGCCTCAA 101 2133 882010 N/A N/A  7304  7319 TAACAAGTTATCCAGT  88 2134 882034 N/A N/A  7593  7608 TGCGAATGTGCCTTGT  57 2135 882058 N/A N/A  8095  8110 CAGCACCGTGTGGAAA  49 2136 882082 N/A N/A  8335  8350 TGGCACCAAAAGTACC  66 2137 882105 N/A N/A  8485  8500 CTTATTAGACTGACAG  59 2138 882129 N/A N/A  9287  9302 CCACATTAGCAATTAG  81 2139 882152 N/A N/A  9550  9565 TTTATGAGCTTCCACA  46 2140 882175 N/A N/A  9843  9858 CCGCATTGCTAGATTC  28 2141 882198 N/A N/A 10115 10130 TTGGAAACCAGCCCTT  61 2142 882222 N/A N/A 10253 10268 TCTACCTACTCCATCA  53 2143 882246 N/A N/A 10580 10595 AGGAAACTTGGAGCGC  23 2144 882270 N/A N/A 11025 11040 GCAAAGCAAGTGTCTA  57 2145 882294 N/A N/A 11313 11328 GAAGGATGATCAGCTT  74 2146 882318 N/A N/A 11536 11551 TTATAGTAGGTAAGAT  98 2147 882342 N/A N/A 11819 11834 TGCAAGCCTCATTCAC  58 2148 882366 N/A N/A 12089 12104 AGTCAGACAGTGCGCC  90 2149 882390 N/A N/A 12380 12395 AAAATATCGCCCATGA  59 2150 882413 N/A N/A 12827 12842 TGACATCATTTAGGTA  46 2151 882436 N/A N/A 13133 13148 GGCCACCGACTCTTTT  87 2152 882459 N/A N/A 13780 13795 ATAACGAGGTGCCTTA  87 2153 882481 N/A N/A 14077 14092 CGCCACATCAGCAGAC  88 2154 882527 N/A N/A 14658 14673 ACTTACACCCAGAAGT 110 2155 882550 N/A N/A 15328 15343 GGCCTTAGCCTTCCTG  83 2156 882574 N/A N/A 15765 15780 GCAGAAGCTGGTTGGC  42 2157 882597 N/A N/A 15877 15892 TGAGACTACACCTACA  81 2158 882620 N/A N/A 16140 16155 ACAGGAGCCGCTACAG  69 2159 882644 N/A N/A 16284 16299 AAGATACAAAACGGAC  61 2160 882667 N/A N/A 16568 16583 CCTCACTACAGAGCCC  76 2161 882691 N/A N/A 16778 16793 TCAAGAGGAACTAGTC  76 2162 882715 N/A N/A 17140 17155 GTAGGAGTGTAACTGA  59 2163 882739 N/A N/A 17407 17422 GGAAATACGACAACTT  37 2164 882763 N/A N/A 17694 17709 TTACGAGAGGGTCTGA  49 2165 882787 N/A N/A 17937 17952 TTTTACCTGGTATTGC 102 2166 882811 N/A N/A 18247 18262 GATCATCAACTTCTTA  53 2167 882834 N/A N/A 18719 18734 TGGAACTCATAGGTGT  38 2168 882857 N/A N/A 19164 19179 CTGCATCAAATCAAGT  53 2169 882881 N/A N/A 19390 19405 TCACTATTAAACAGCA  84 2170 882910 N/A N/A 14263 14278 GTTCATATCACAGAGC  55 2171

TABLE 31 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  23  195 690526 4234 4249 22351 22366 GGTTTCAGTTGTAAAT  42 2172 881093  193  208  5220  5235 GCCACTGGCGGAGCTT  77 2173 881116  554  569  7872  7887 ATGGACATCTGCGGGT  50 2174 881163 1090 1105 13643 13658 TCGCATAGAGCCCGTC  43 2175 881187 1274 1289 17067 17082 CCAAAGCATAGAGTCA  42 2176 881211 1539 1554 19656 19671 CAAGACCCCGTATCCC  56 2177 881235 1897 1912 20014 20029 CAATGATGGCAGTCTA  60 2178 881259 2015 2030 20132 20147 TGTCAATACTGAAAGG  64 2179 881283 2156 2171 20273 20288 CTCAGCAAGTGTCTTC  55 2180 881307 2317 2332 20434 20449 ATAGGAAGTTTACTAA  87 2181 881331 2465 2480 20582 20597 ATGCGAAGAATTTTAG  79 2182 881355 2583 2598 20700 20715 GGACAGCCAAACAAAC  93 2183 881379 2778 2793 20895 20910 CAAGTAATTTTAAGGC  59 2184 881403 2849 2864 20966 20981 TGTAAGTGATAGGTAT  47 2185 881427 2973 2988 21090 21105 ACACATTTGAGAGCTA  31 2186 881451 3254 3269 21371 21386 GGACACTTTTAGAGAG  38 2187 881475 3397 3412 21514 21529 AGAACGAGGTAAATCT  60 2188 881499 3486 3501 21603 21618 GCCCAATACATATCCA  42 2189 881522 3601 3616 21718 21733 CCGTGAAACAGCAGTT  44 2190 881546 3791 3806 21908 21923 AGCTCCACTGTTAAAG  56 2191 881570 3999 4014 22116 22131 CTTTATCAAGACAAGG  44 2192 881613 4388 4403 22505 22520 TAACAATACTGTAAGG  55 2193 881637 4523 4538 22640 22655 GCGGAGCATCAACAAA  70 2194 881661 4594 4609 22711 22726 ACGGAAGTTTACACTG  23 2195 881685 4810 4825 22927 22942 GCATCGAGATCAGTCT  43 2196 881709 4994 5009 23111 23126 AAGCAACACGGGAATT  78 2197 881733 5198 5213 23315 23330 GCATTATCTACCTCAG  31 2198 881757 N/A N/A 18769 18784 AGCTAAACTCCTAAGT  85 2199 881781 N/A N/A 18832 18847 GGCAAGCTGAATGCTC  71 2200 881804 N/A N/A  4002  4017 AAGGAGGCGGGCCTTG  98 2201 881826 N/A N/A  4315  4330 CCGCACGCAAGGCTCG  84 2202 881849 N/A N/A  4563  4578 TCGCATCCAGACCCTT  72 2203 881873 N/A N/A  4939  4954 CGGAAGAAGGCACCTT  112 2204 881897 N/A N/A  5647  5662 CGCAAAACCCCTCAAG  81 2205 881920 N/A N/A  5760  5775 CACAGGCGCGGACGCA  86 2206 881941 N/A N/A  6021  6036 GTAACAACGACACACG  41 2207 881965 N/A N/A  6325  6340 CCCTATCACTGCCAAG  62 2208 881987 N/A N/A  7065  7080 CAGAATGAAGTTAAGG  50 2209 882011 N/A N/A  7325  7340 GACTATTAGATAAGTA  71 2210 882035 N/A N/A  7602  7617 CAGATGGCATGCGAAT  65 2211 882059 N/A N/A  8097  8112 GGCAGCACCGTGTGGA  47 2212 882083 N/A N/A  8344  8359 GGCTAAACCTGGCACC  60 2213 882106 N/A N/A  8486  8501 CCTTATTAGACTGACA  52 2214 882130 N/A N/A  9288  9303 GCCACATTAGCAATTA  92 2215 882153 N/A N/A  9557  9572 CTTATTATTTATGAGC  63 2216 882176 N/A N/A  9845  9860 TACCGCATTGCTAGAT  77 2217 882199 N/A N/A 10119 10134 CATATTGGAAACCAGC  27 2218 882223 N/A N/A 10264 10279 CAAGACAGGTCTCTAC  66 2219 882247 N/A N/A 10581 10596 AAGGAAACTTGGAGCG  41 2220 882271 N/A N/A 11026 11041 AGCAAAGCAAGTGTCT  42 2221 882295 N/A N/A 11321 11336 TCAACCTGGAAGGATG  70 2222 882319 N/A N/A 11537 11552 TTTATAGTAGGTAAGA  94 2223 882343 N/A N/A 11827 11842 AAAAAAGGTGCAAGCC  93 2224 882367 N/A N/A 12093 12108 AGCGAGTCAGACAGTG  84 2225 882391 N/A N/A 12382 12397 TTAAAATATCGCCCAT  68 2226 882414 N/A N/A 12871 12886 AGCAAGTCGGTCCACA  46 2227 882437 N/A N/A 13154 13169 CAGCATATTACAAACG  42 2228 882460 N/A N/A 13781 13796 GATAACGAGGTGCCTT  82 2229 882504 N/A N/A 14295 14310 ACTCACTGGTTCTGAA  86 2230 882528 N/A N/A 14661 14676 GTCACTTACACCCAGA  48 2231 882551 N/A N/A 15359 15374 GTCCAGAGTCTTCAGA  94 2232 882575 N/A N/A 15772 15787 TAATTATGCAGAAGCT 103 2233 882598 N/A N/A 15879 15894 TCTGAGACTACACCTA  73 2234 882621 N/A N/A 16146 16161 GTTGACACAGGAGCCG  47 2235 882645 N/A N/A 16285 16300 AAAGATACAAAACGGA  62 2236 882668 N/A N/A 16581 16596 TAAAACCTCATCCCCT 126 2237 882692 N/A N/A 16779 16794 TTCAAGAGGAACTAGT  94 2238 882716 N/A N/A 17146 17161 ACTATGGTAGGAGTGT  75 2239 882740 N/A N/A 17409 17424 ATGGAAATACGACAAC  65 2240 882764 N/A N/A 17696 17711 CATTACGAGAGGGTCT  56 2241 882788 N/A N/A 17940 17955 AACTTTTACCTGGTAT  72 2242 882812 N/A N/A 18251 18266 GGCCGATCATCAACTT  93 2243 882835 N/A N/A 18720 18735 GTGGAACTCATAGGTG  47 2244 882858 N/A N/A 19173 19188 TAACTAAAACTGCATC  90 2245 882882 N/A N/A 19391 19406 CTCACTATTAAACAGC  76 2246 882909 N/A N/A 14080 14095 AGCCGCCACATCAGCA  79 2247

TABLE 32 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  26  195 881094  194  209  5221  5236 AGCCACTGGCGGAGCT  62 2248 881117  558  573  7876  7891 GCTCATGGACATCTGC  61 2249 881164 1097 1112 13650 13665 AGTCTTTTCGCATAGA  55 2250 881188 1276 1291 17069 17084 CTCCAAAGCATAGAGT  77 2251 881212 1685 1700 19802 19817 AATCACTTGTCTTGGG  40 2252 881236 1898 1913 20015 20030 TCAATGATGGCAGTCT  46 2253 881260 2021 2036 20138 20153 AGTCAGTGTCAATACT  53 2254 881284 2157 2172 20274 20289 ACTCAGCAAGTGTCTT  53 2255 881308 2341 2356 20458 20473 GCTCATGTTTTTTGAC  44 2256 881332 2469 2484 20586 20601 CTTTATGCGAAGAATT  70 2257 881356 2587 2602 20704 20719 CGCTGGACAGCCAAAC  59 2258 881380 2780 2795 20897 20912 GCCAAGTAATTTTAAG  65 2259 881404 2850 2865 20967 20982 ATGTAAGTGATAGGTA  30 2260 881428 2995 3010 21112 21127 TATCCATTAGAAAAGC  37 2261 881452 3255 3270 21372 21387 TGGACACTTTTAGAGA  28 2262 881476 3398 3413 21515 21530 CAGAACGAGGTAAATC  41 2263 881500 3487 3502 21604 21619 TGCCCAATACATATCC  41 2264 881523 3611 3626 21728 21743 GGTAAGGGCCCCGTGA  67 2265 881547 3835 3850 21952 21967 AAGGAGGTAGTGGCCC  53 2266 881571 4009 4024 22126 22141 GCCAATTCCACTTTAT  36 2267 881590 4239 4254 22356 22371 TCCTAGGTTTCAGTTG  52 2268 881614 4389 4404 22506 22521 GTAACAATACTGTAAG  45 2269 881638 4524 4539 22641 22656 GGCGGAGCATCAACAA  40 2270 881662 4595 4610 22712 22727 AACGGAAGTTTACACT  41 2271 881686 4812 4827 22929 22944 CTGCATCGAGATCAGT  48 2272 881710 4996 5011 23113 23128 TGAAGCAACACGGGAA  40 2273 881734 5199 5214 23316 23331 AGCATTATCTACCTCA  22 2274 881758 N/A N/A 18770 18785 AAGCTAAACTCCTAAG  85 2275 881782 N/A N/A 18837 18852 TCTAAGGCAAGCTGAA  79 2276 881805 N/A N/A  4003  4018 CAAGGAGGCGGGCCTT 102 2277 881827 N/A N/A  4325  4340 GGCTAGGCCACCGCAC  72 2278 881850 N/A N/A  4633  4648 GAGGACCTCGCCTGCG 102 2279 881874 N/A N/A  4940  4955 CCGGAAGAAGGCACCT  83 2280 881898 N/A N/A  5648  5663 GCGCAAAACCCCTCAA  78 2281 881921 N/A N/A  5763  5778 CGGCACAGGCGCGGAC  96 2282 881942 N/A N/A  6047  6062 GTAAACTGCCTTAGGG  56 2283 881966 N/A N/A  6369  6384 AAAGACCCCAGCTATG  77 2284 881988 N/A N/A  7068  7083 GCTCAGAATGAAGTTA  52 2285 882012 N/A N/A  7327  7342 AAGACTATTAGATAAG  78 2286 882036 N/A N/A  7629  7644 GTACAGGACAGGTAAA  75 2287 882060 N/A N/A  8103  8118 ACCAATGGCAGCACCG  39 2288 882084 N/A N/A  8347  8362 TATGGCTAAACCTGGC  57 2289 882107 N/A N/A  8487  8502 CCCTTATTAGACTGAC  33 2290 882131 N/A N/A  9312  9327 GACCAGGATTCGCCAT  77 2291 882177 N/A N/A  9850  9865 TGAGTTACCGCATTGC  20 2292 882200 N/A N/A 10121 10136 ACCATATTGGAAACCA  26 2293 882224 N/A N/A 10278 10293 TGTTACCGATGCTTCA  51 2294 882248 N/A N/A 10606 10621 GAAACGAGCCAGTGCA  48 2295 882272 N/A N/A 11027 11042 GAGCAAAGCAAGTGTC  58 2296 882296 N/A N/A 11323 11338 GCTCAACCTGGAAGGA  75 2297 882320 N/A N/A 11541 11556 CTTCTTTATAGTAGGT  80 2298 882344 N/A N/A 11843 11858 TCACGAGCACCTCAGA  64 2299 882368 N/A N/A 12111 12126 TTCAACACACTGTCTG 131 2300 882392 N/A N/A 12386 12401 CTCTTTAAAATATCGC  44 2301 882415 N/A N/A 12872 12887 AAGCAAGTCGGTCCAC  68 2302 882438 N/A N/A 13161 13176 TCTAAACCAGCATATT  67 2303 882482 N/A N/A 14085 14100 AAAAGAGCCGCCACAT  84 2304 882505 N/A N/A 14321 14336 CATGAAATCCAAGGTA  51 2305 882529 N/A N/A 14671 14686 AAAAACTTGGGTCACT 119 2306 882552 N/A N/A 15366 15381 CACCAAGGTCCAGAGT  80 2307 882599 N/A N/A 15893 15908 TCACACTGCCGTGATC  78 2308 882622 N/A N/A 16151 16166 AACGAGTTGACACAGG  45 2309 882646 N/A N/A 16314 16329 GTACATCCTGAAGCCA  74 2310 882669 N/A N/A 16612 16627 ATCAGAATGTTTCGAC  54 2311 882693 N/A N/A 16807 16822 CCCCAGGGCCCTCGGT  82 2312 882717 N/A N/A 17147 17162 CACTATGGTAGGAGTG 112 2313 882741 N/A N/A 17448 17463 GGGCAACTTTAACCAT  92 2314 882765 N/A N/A 17699 17714 GAACATTACGAGAGGG  30 2315 882789 N/A N/A 17945 17960 GGTGTAACTTTTACCT  89 2316 882813 N/A N/A 18558 18573 GATCATCAACTTCTTT  57 2317 882836 N/A N/A 18760 18775 CCTAAGTACCTGAAAT  89 2318 882859 N/A N/A 19203 19218 TCAAATCGACTGCCAC  46 2319 882883 N/A N/A 19392 19407 GCTCACTATTAAACAG 112 2320 882902 N/A N/A  9579  9594 CAATAATCTCCCAGCA  63 2321 882907 N/A N/A 13782 13797 AGATAACGAGGTGCCT  77 2322 882912 N/A N/A 15773 15788 TTAATTATGCAGAAGC  87 2323

TABLE 33 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  26  195 881095  217  232  5244  5259 TGCCGCTGTCGATCTG  68 2324 881118  559  574  7877  7892 GGCTCATGGACATCTG  64 2325 881165 1103 1118 13656 13671 TGGCACAGTCTTTTCG  72 2326 881189 1341 1356 19458 19473 GGCTAGCAGAGGTTCT  49 2327 881213 1717 1732 19834 19849 CGCTTTCACTAAAGTC  29 2328 881237 1900 1915 20017 20032 CATCAATGATGGCAGT  50 2329 881261 2025 2040 20142 20157 CTCTAGTCAGTGTCAA  41 2330 881285 2158 2173 20275 20290 CACTCAGCAAGTGTCT  50 2331 881309 2358 2373 20475 20490 TCCCATCCAAGAGTAG  67 2332 881333 2470 2485 20587 20602 TCTTTATGCGAAGAAT  58 2333 881357 2600 2615 20717 20732 CGCCATGGCTGATCGC  47 2334 881381 2817 2832 20934 20949 GTCTTTGGGATTCTAT  35 2335 881405 2851 2866 20968 20983 AATGTAAGTGATAGGT  20 2336 881429 3024 3039 21141 21156 TACTAGGTGCTTGTTG  51 2337 881453 3256 3271 21373 21388 GTGGACACTTTTAGAG  48 2338 881477 3400 3415 21517 21532 AGCAGAACGAGGTAAA  30 2339 881501 3500 3515 21617 21632 AAACACAGTCTGCTGC  65 2340 881524 3612 3627 21729 21744 AGGTAAGGGCCCCGTG  53 2341 881548 3836 3851 21953 21968 AAAGGAGGTAGTGGCC  79 2342 881572 4022 4037 22139 22154 TAAATTCTAGTTTGCC  32 2343 881591 4258 4273 22375 22390 CGCTCAGGACTCAGGG  45 2344 881615 4390 4405 22507 22522 GGTAACAATACTGTAA  41 2345 881639 4525 4540 22642 22657 TGGCGGAGCATCAACA  63 2346 881663 4596 4611 22713 22728 GAACGGAAGTTTACAC  33 2347 881687 4817 4832 22934 22949 TCCACCTGCATCGAGA  53 2348 881711 4997 5012 23114 23129 TTGAAGCAACACGGGA  39 2349 881735 5202 5217 23319 23334 CATAGCATTATCTACC  57 2350 881759 N/A N/A 18772 18787 TTAAGCTAAACTCCTA  90 2351 881783 N/A N/A 18840 18855 GCATCTAAGGCAAGCT  66 2352 881806 N/A N/A  4006  4021 AGCCAAGGAGGCGGGC 101 2353 881828 N/A N/A  4334  4349 CGCCAGGCCGGCTAGG  90 2354 881851 N/A N/A  4636  4651 GCGGAGGACCTCGCCT  92 2355 881875 N/A N/A  4942  4957 CCCCGGAAGAAGGCAC  64 2356 881899 N/A N/A  5654  5669 ACGAACGCGCAAAACC  94 2357 881922 N/A N/A  5769  5784 AGCCGCCGGCACAGGC  99 2358 881943 N/A N/A  6048  6063 GGTAAACTGCCTTAGG  75 2359 881967 N/A N/A  6371  6386 TGAAAGACCCCAGCTA  96 2360 881989 N/A N/A  7080  7095 CTAGAAAGTGATGCTC  61 2361 882013 N/A N/A  7332  7347 CACTAAAGACTATTAG  95 2362 882037 N/A N/A  7631  7646 TTGTACAGGACAGGTA  72 2363 882061 N/A N/A  8109  8124 ATCCACACCAATGGCA  66 2364 882108 N/A N/A  8494  8509 AGGGAATCCCTTATTA  94 2365 882132 N/A N/A  9317  9332 GGACAGACCAGGATTC  75 2366 882154 N/A N/A  9580  9595 TCAATAATCTCCCAGC  45 2367 882178 N/A N/A  9853  9868 GCCTGAGTTACCGCAT  30 2368 882201 N/A N/A 10122 10137 TACCATATTGGAAACC  52 2369 882225 N/A N/A 10283 10298 TTTACTGTTACCGATG  52 2370 882249 N/A N/A 10608 10623 GAGAAACGAGCCAGTG  61 2371 882273 N/A N/A 11048 11063 ATCTACTCCAGACCCC  73 2372 882297 N/A N/A 11324 11339 GGCTCAACCTGGAAGG  71 2373 882321 N/A N/A 11552 11567 CCTAGAGGTGCCTTCT  59 2374 882345 N/A N/A 11846 11861 TACTCACGAGCACCTC  59 2375 882369 N/A N/A 12114 12129 TGCTTCAACACACTGT  97 2376 882393 N/A N/A 12432 12447 TAAACAAGATGAATCC  56 2377 882416 N/A N/A 12875 12890 AAGAAGCAAGTCGGTC  45 2378 882439 N/A N/A 13162 13177 GTCTAAACCAGCATAT  81 2379 882461 N/A N/A 13783 13798 CAGATAACGAGGTGCC  70 2380 882483 N/A N/A 14086 14101 GAAAAGAGCCGCCACA  93 2381 882506 N/A N/A 14331 14346 GCTGAATTGTCATGAA  64 2382 882553 N/A N/A 15369 15384 TGGCACCAAGGTCCAG  99 2383 882576 N/A N/A 15776 15791 TCCTTAATTATGCAGA  56 2384 882600 N/A N/A 15895 15910 ATTCACACTGCCGTGA  66 2385 882623 N/A N/A 16152 16167 AAACGAGTTGACACAG  80 2386 882670 N/A N/A 16646 16661 CAATTTATGCCATGGA  38 2387 882694 N/A N/A 16838 16853 TAGCATGTATGCATTC  48 2388 882718 N/A N/A 17150 17165 AGCCACTATGGTAGGA  57 2389 882742 N/A N/A 17471 17486 ATCTTAACCTGGAGAA  72 2390 882766 N/A N/A 17703 17718 TAGAGAACATTACGAG  40 2391 882790 N/A N/A 17976 17991 CTAGAACATGATGAGA  71 2392 882837 N/A N/A 18921 18936 TTATACTGTCTGGTTA  56 2393 882860 N/A N/A 19205 19220 TTTCAAATCGACTGCC  69 2394 882884 N/A N/A 19400 19415 TGCAACTGGCTCACTA  83 2395 882900 N/A N/A  8350  8365 TCATATGGCTAAACCT  51 2396 882911 N/A N/A 14672 14687 CAAAAACTTGGGTCAC  80 2397 882914 N/A N/A 16378 16393 GTAACTTGACTTGAGA  51 2398 882915 N/A N/A 18567 18582 AGCTGAGCTGATCATC  44 2399

TABLE 34 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  24  195 881096  328  343  5355  5370 CCTTGAAGAGCGCGGC  76 2400 881119  594  609  7912  7927 GAGCGAAGGGTAAGGC  62 2401 881166 1121 1136 13674 13689 TCCCAGTAGATCCTGC  60 2402 881190 1354 1369 19471 19486 AATATAGTTGTCTGGC  50 2403 881214 1732 1747 19849 19864 GGGCAGTCAATTGGAC  75 2404 881238 1903 1918 20020 20035 GATCATCAATGATGGC  37 2405 881262 2033 2048 20150 20165 AGTCATCACTCTAGTC  47 2406 881286 2189 2204 20306 20321 GGCACGGCTTCAGTCA  25 2407 881310 2366 2381 20483 20498 CAAAAATGTCCCATCC  81 2408 881334 2471 2486 20588 20603 TTCTTTATGCGAAGAA  91 2409 881358 2609 2624 20726 20741 CTTTAGTGTCGCCATG  71 2410 881382 2822 2837 20939 20954 TGGAGGTCTTTGGGAT  42 2411 881406 2882 2897 20999 21014 GTCTACTGCTGTACTT  48 2412 881430 3025 3040 21142 21157 TTACTAGGTGCTTGTT  50 2413 881454 3258 3273 21375 21390 TTGTGGACACTTTTAG  51 2414 881478 3401 3416 21518 21533 GAGCAGAACGAGGTAA  45 2415 881502 3502 3517 21619 21634 CGAAACACAGTCTGCT  47 2416 881525 3616 3631 21733 21748 TCACAGGTAAGGGCCC  53 2417 881549 3837 3852 21954 21969 GAAAGGAGGTAGTGGC  77 2418 881573 4023 4038 22140 22155 CTAAATTCTAGTTTGC  79 2419 881592 4276 4291 22393 22408 GACTAACTCTCCTGTT  83 2420 881616 4391 4406 22508 22523 TGGTAACAATACTGTA  35 2421 881640 4533 4548 22650 22665 GGCCTTCCTGGCGGAG  78 2422 881664 4598 4613 22715 22730 ATGAACGGAAGTTTAC  46 2423 881688 4828 4843 22945 22960 TCTCAAGGAGATCCAC  99 2424 881712 4998 5013 23115 23130 TTTGAAGCAACACGGG  53 2425 881736 5203 5218 23320 23335 GCATAGCATTATCTAC  49 2426 881760 N/A N/A 18774 18789 ACTTAAGCTAAACTCC  79 2427 881784 N/A N/A 18841 18856 AGCATCTAAGGCAAGC  43 2428 881807 N/A N/A  4042  4057 CAGGAGCGCGGAGGGC  89 2429 881829 N/A N/A  4367  4382 ACGACAGCTGCGGAGC  73 2430 881852 N/A N/A  4638  4653 GCGCGGAGGACCTCGC  90 2431 881876 N/A N/A  5008  5023 GACCACGAGGCCCCGG  69 2432 881900 N/A N/A  5656  5671 GGACGAACGCGCAAAA  65 2433 881923 N/A N/A  5773  5788 AAACAGCCGCCGGCAC  89 2434 881944 N/A N/A  6117  6132 TCCCGGAGGAAGTCCC  70 2435 881968 N/A N/A  6377  6392 GTAAACTGAAAGACCC  65 2436 881990 N/A N/A  7152  7167 CATAACTCAGGCAAGG  64 2437 882014 N/A N/A  7335  7350 ACTCACTAAAGACTAT  82 2438 882038 N/A N/A  7645  7660 CTACAAGGTCTGAGTT  83 2439 882062 N/A N/A  8116  8131 ACTTAAAATCCACACC  93 2440 882085 N/A N/A  8366  8381 TTTATGTAAAGCTTCG  25 2441 882109 N/A N/A  8498  8513 CCTCAGGGAATCCCTT  70 2442 882133 N/A N/A  9340  9355 GGAAAGAGCTTTGGTG  88 2443 882155 N/A N/A  9581  9596 CTCAATAATCTCCCAG  47 2444 882179 N/A N/A  9877  9892 GTAACTAACTCAAAAG 101 2445 882202 N/A N/A 10125 10140 CACTACCATATTGGAA  78 2446 882226 N/A N/A 10284 10299 ATTTACTGTTACCGAT  71 2447 882250 N/A N/A 10609 10624 GGAGAAACGAGCCAGT  53 2448 882274 N/A N/A 11052 11067 CTACATCTACTCCAGA  78 2449 882298 N/A N/A 11332 11347 TTATTTGTGGCTCAAC  82 2450 882322 N/A N/A 11554 11569 AGCCTAGAGGTGCCTT  59 2451 882346 N/A N/A 11859 11874 GGGCATCCTCAGTTAC  67 2452 882370 N/A N/A 12138 12153 ACCCACATCACTGTCT  85 2453 882394 N/A N/A 12456 12471 CTAAAACTGCGCTCTC  76 2454 882417 N/A N/A 12878 12893 AGAAAGAAGCAAGTCG  63 2455 882440 N/A N/A 13169 13184 TTAATCTGTCTAAACC  84 2456 882462 N/A N/A 13784 13799 ACAGATAACGAGGTGC  70 2457 882484 N/A N/A 14116 14131 TTCCAACCTTTATGAT  93 2458 882507 N/A N/A 14335 14350 GTTCGCTGAATTGTCA  63 2459 882530 N/A N/A 14674 14689 TACAAAAACTTGGGTC  91 2460 882554 N/A N/A 15406 15421 TAGAAAGCCCTCACCT  95 2461 882577 N/A N/A 15781 15796 GGGAATCCTTAATTAT  88 2462 882601 N/A N/A 15898 15913 TACATTCACACTGCCG  42 2463 882624 N/A N/A 16154 16169 AGAAACGAGTTGACAC  85 2464 882647 N/A N/A 16380 16395 CAGTAACTTGACTTGA  61 2465 882671 N/A N/A 16647 16662 CCAATTTATGCCATGG  88 2466 882695 N/A N/A 16857 16872 GAACATCTCAGATACA  58 2467 882719 N/A N/A 17158 17173 GAACAGGAAGCCACTA  77 2468 882743 N/A N/A 17481 17496 TCTTACAGCAATCTTA  68 2469 882767 N/A N/A 17709 17724 CTATGCTAGAGAACAT  77 2470 882791 N/A N/A 17977 17992 GCTAGAACATGATGAG  86 2471 882814 N/A N/A 18570 18585 GATAGCTGAGCTGATC  68 2472 882838 N/A N/A 18923 18938 ATTTATACTGTCTGGT  51 2473 882861 N/A N/A 19211 19226 TGTTACTTTCAAATCG  52 2474 882885 N/A N/A 19401 19416 CTGCAACTGGCTCACT  90 2475

TABLE 35 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  12  195 881097  329  344 N/A N/A GCCTTGAAGAGCGCGG  91 2476 881120  613  628 N/A N/A TGTGAACCTGCTGGGC  98 2477 881167 1189 1204 13742 13757 CAAAGAGCTTGCAGGT  96 2478 881191 1355 1370 19472 19487 TAATATAGTTGTCTGG  60 2479 881215 1738 1753 19855 19870 GTAAGAGGGCAGTCAA  56 2480 881239 1910 1925 20027 20042 TCACAGTGATCATCAA  57 2481 881263 2040 2055 20157 20172 TACAAGCAGTCATCAC  71 2482 881287 2190 2205 20307 20322 AGGCACGGCTTCAGTC  51 2483 881311 2382 2397 20499 20514 TAGATTGTAGGACAGA  72 2484 881335 2472 2487 20589 20604 CTTCTTTATGCGAAGA 105 2485 881359 2610 2625 20727 20742 CCTTTAGTGTCGCCAT  42 2486 881383 2824 2839 20941 20956 AGTGGAGGTCTTTGGG  45 2487 881407 2887 2902 21004 21019 CCCCAGTCTACTGCTG  53 2488 881431 3026 3041 21143 21158 CTTACTAGGTGCTTGT  62 2489 881455 3270 3285 21387 21402 AAACACCCCTTCTTGT 113 2490 881479 3404 3419 21521 21536 TCTGAGCAGAACGAGG  53 2491 881503 3503 3518 21620 21635 ACGAAACACAGTCTGC  48 2492 881526 3618 3633 21735 21750 GGTCACAGGTAAGGGC  75 2493 881550 3896 3911 22013 22028 TCATACCCTGGATCAC  57 2494 881574 4039 4054 22156 22171 GTCCACTGAGTACAAA  87 2495 881593 4278 4293 22395 22410 GCGACTAACTCTCCTG  30 2496 881617 4410 4425 22527 22542 AATACCTACCTGCCCT  85 2497 881641 4540 4555 22657 22672 CACAAGTGGCCTTCCT  55 2498 881665 4600 4615 22717 22732 CAATGAACGGAAGTTT  67 2499 881689 4837 4852 22954 22969 CTATCAGGATCTCAAG  91 2500 881713 5020 5035 23137 23152 TGTTAAGTCCCATCTG  75 2501 881737 5204 5219 23321 23336 AGCATAGCATTATCTA  66 2502 881761 N/A N/A 18775 18790 GACTTAAGCTAAACTC  42 2503 881785 N/A N/A 18842 18857 CAGCATCTAAGGCAAG  46 2504 881830 N/A N/A  4368  4383 GACGACAGCTGCGGAG  55 2505 881853 N/A N/A  4640  4655 ACGCGCGGAGGACCTC  87 2506 881877 N/A N/A  5011  5026 AGTGACCACGAGGCCC  85 2507 881901 N/A N/A  5657  5672 CGGACGAACGCGCAAA  61 2508 881924 N/A N/A  5775  5790 GAAAACAGCCGCCGGC  74 2509 881945 N/A N/A  6122  6137 CAAATTCCCGGAGGAA  95 2510 881969 N/A N/A  6378  6393 CGTAAACTGAAAGACC  52 2511 881991 N/A N/A  7153  7168 ACATAACTCAGGCAAG  99 2512 882015 N/A N/A  7345  7360 CAAAATAGTCACTCAC 103 2513 882039 N/A N/A  7646  7661 TCTACAAGGTCTGAGT  43 2514 882063 N/A N/A  8119  8134 CCAACTTAAAATCCAC  91 2515 882086 N/A N/A  8380  8395 GATACTTGTACTGTTT  22 2516 882110 N/A N/A  8546  8561 AAGAAGCACTGGCATT  78 2517  9066  9081 882134 N/A N/A  9341  9356 GGGAAAGAGCTTTGGT  95 2518 882156 N/A N/A  9592  9607 AAATATACCAGCTCAA  75 2519 882180 N/A N/A  9921  9936 CCAAAGGGTTCAGTGT  74 2520 882203 N/A N/A 10128 10143 CTCCACTACCATATTG  99 2521 882227 N/A N/A 10285 10300 CATTTACTGTTACCGA  21 2522 882251 N/A N/A 10610 10625 TGGAGAAACGAGCCAG  62 2523 882275 N/A N/A 11054 11069 GTCTACATCTACTCCA  62 2524 882299 N/A N/A 11333 11348 CTTATTTGTGGCTCAA  45 2525 882323 N/A N/A 11655 11670 ACCTTAAGCTATTTGG  39 2526 882347 N/A N/A 11865 11880 CCCAAAGGGCATCCTC  59 2527 882371 N/A N/A 12143 12158 ACTTAACCCACATCAC 122 2528 882395 N/A N/A 12484 12499 GGCTTTGTGTTTAAGT  77 2529 882418 N/A N/A 12890 12905 TAACGGTGTTTCAGAA  74 2530 882441 N/A N/A 13173 13188 CCATTTAATCTGTCTA  72 2531 882463 N/A N/A 13785 13800 AACAGATAACGAGGTG 104 2532 882485 N/A N/A 14124 14139 CAATATGCTTCCAACC 115 2533 882508 N/A N/A 14343 14358 AGCCAGAGGTTCGCTG  75 2534 882531 N/A N/A 14677 14692 GCTTACAAAAACTTGG  74 2535 882555 N/A N/A 15407 15422 TTAGAAAGCCCTCACC 116 2536 882578 N/A N/A 15782 15797 TGGGAATCCTTAATTA  88 2537 882625 N/A N/A 16155 16170 AAGAAACGAGTTGACA  78 2538 882648 N/A N/A 16385 16400 TGGAACAGTAACTTGA  88 2539 882672 N/A N/A 16648 16663 ACCAATTTATGCCATG  83 2540 882696 N/A N/A 16860 16875 TGTGAACATCTCAGAT  88 2541 882720 N/A N/A 17170 17185 GGCCTTTACAAAGAAC 119 2542 882744 N/A N/A 17487 17502 TAGCATTCTTACAGCA  43 2543 882768 N/A N/A 17726 17741 GTTGAACCCATCTTGA  84 2544 882792 N/A N/A 17978 17993 AGCTAGAACATGATGA  95 2545 882815 N/A N/A 18571 18586 TGATAGCTGAGCTGAT  63 2546 882839 N/A N/A 18939 18954 AGCTAACTGGCCTCAA  77 2547 882862 N/A N/A 19221 19236 GGGTTAAAGATGTTAC  50 2548 882886 N/A N/A 19408 19423 AGATATCCTGCAACTG 109 2549 882891 N/A N/A  4045  4060 TCGCAGGAGCGCGGAG 137 2550 882913 N/A N/A 15899 15914 ATACATTCACACTGCC  55 2551

TABLE 36 Percent control of human IRF4 mRNA with 3-10-3 cEt gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ Number Site Site Site Site Sequence UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  26  195 881098  335  350 N/A N/A GCCCAAGCCTTGAAGA  85 2552 881121  620  635  9110  9125 ATGTAGTTGTGAACCT  61 2553 881168 1191 1206 13744 13759 GTCAAAGAGCTTGCAG  56 2554 881192 1356 1371 19473 19488 ATAATATAGTTGTCTG  65 2555 881216 1739 1754 19856 19871 AGTAAGAGGGCAGTCA  50 2556 881240 1919 1934 20036 20051 GGTCAATTTTCACAGT  36 2557 881264 2041 2056 20158 20173 CTACAAGCAGTCATCA  56 2558 881288 2192 2207 20309 20324 ACAGGCACGGCTTCAG  36 2559 881312 2384 2399 20501 20516 ACTAGATTGTAGGACA 109 2560 881336 2507 2522 20624 20639 GTTCAAGTATTAAGAT  71 2561 881360 2611 2626 20728 20743 TCCTTTAGTGTCGCCA  24 2562 881384 2825 2840 20942 20957 AAGTGGAGGTCTTTGG  34 2563 881432 3043 3058 21160 21175 TAGGGATACAGCAGGC  27 2564 881456 3272 3287 21389 21404 ATAAACACCCCTTCTT  70 2565 881480 3408 3423 21525 21540 GGCCTCTGAGCAGAAC  73 2566 881504 3504 3519 21621 21636 CACGAAACACAGTCTG  48 2567 881527 3624 3639 21741 21756 AAAGAGGGTCACAGGT  66 2568 881551 3898 3913 22015 22030 GGTCATACCCTGGATC  51 2569 881575 4051 4066 22168 22183 TTCAACAGCACTGTCC  35 2570 881594 4284 4299 22401 22416 TGTAGGGCGACTAACT  79 2571 881618 4411 4426 22528 22543 TAATACCTACCTGCCC  96 2572 881642 4543 4558 22660 22675 ACACACAAGTGGCCTT  56 2573 881666 4601 4616 22718 22733 GCAATGAACGGAAGTT  55 2574 881690 4838 4853 22955 22970 GCTATCAGGATCTCAA  62 2575 881714 5021 5036 23138 23153 CTGTTAAGTCCCATCT  46 2576 881738 5205 5220 23322 23337 CAGCATAGCATTATCT 151 2577 881762 N/A N/A 18777 18792 ACGACTTAAGCTAAAC  42 2578 881786 N/A N/A 18846 18861 TTTACAGCATCTAAGG  66 2579 881808 N/A N/A  4055  4070 GGCGACCCCGTCGCAG  70 2580 881831 N/A N/A  4372  4387 GGGCGACGACAGCTGC  68 2581 881854 N/A N/A  4642  4657 CCACGCGCGGAGGACC  81 2582 881878 N/A N/A  5015  5030 CGCCAGTGACCACGAG  58 2583 881902 N/A N/A  5659  5674 CACGGACGAACGCGCA 102 2584 881946 N/A N/A  6124  6139 ACCAAATTCCCGGAGG  93 2585 881970 N/A N/A  6413  6428 GCATAGGTCCTTCAGA  69 2586 881992 N/A N/A  7154  7169 CACATAACTCAGGCAA  63 2587 882016 N/A N/A  7347  7362 TGCAAAATAGTCACTC  52 2588 882040 N/A N/A  7647  7662 TTCTACAAGGTCTGAG  53 2589 882064 N/A N/A  8139  8154 GCGGACACGCCCGACC  85 2590 882087 N/A N/A  8383  8398 GGAGATACTTGTACTG  34 2591 882111 N/A N/A  8550  8565 AGATAAGAAGCACTGG  75 2592  8706  8721  8810  8825  8966  8981  9070  9085 882157 N/A N/A  9593  9608 GAAATATACCAGCTCA  39 2593 882181 N/A N/A  9957  9972 GGCCAAATTGCAAAGG  52 2594 882204 N/A N/A 10149 10164 TAGTTTTATGTTAGCC  25 2595 882228 N/A N/A 10287 10302 TTCATTTACTGTTACC  29 2596 882252 N/A N/A 10615 10630 GCTGATGGAGAAACGA  93 2597 882276 N/A N/A 11087 11102 TAAATCACCCTGGTCA  78 2598 882300 N/A N/A 11347 11362 GTAGAGGAGGAGGACT  84 2599 882324 N/A N/A 11660 11675 TCCAAACCTTAAGCTA  63 2600 882348 N/A N/A 11891 11906 AGCCTTCCTGCTCAGA  45 2601 882372 N/A N/A 12144 12159 GACTTAACCCACATCA 266 2602 882396 N/A N/A 12492 12507 TCCCACTGGGCTTTGT  68 2603 882419 N/A N/A 12891 12906 ATAACGGTGTTTCAGA  53 2604 882442 N/A N/A 13174 13189 CCCATTTAATCTGTCT  67 2605 882464 N/A N/A 13787 13802 CTAACAGATAACGAGG  77 2606 882486 N/A N/A 14129 14144 GTTAACAATATGCTTC  75 2607 882509 N/A N/A 14347 14362 AGCCAGCCAGAGGTTC  70 2608 882532 N/A N/A 14691 14706 GAAAATCTGGATGAGC  36 2609 882556 N/A N/A 15437 15452 AGCCAGTGCCAGTTCC  61 2610 882579 N/A N/A 15803 15818 AGATAACATGAGAGTG  59 2611 882602 N/A N/A 15923 15938 AATGACTTAGTCAGAA  73 2612 882626 N/A N/A 16159 16174 GCAAAAGAAACGAGTT  92 2613 882649 N/A N/A 16387 16402 GATGGAACAGTAACTT  49 2614 882673 N/A N/A 16663 16678 ACGCAATGGCAAAAGA  89 2615 882697 N/A N/A 16887 16902 TCTTACTCCGCTGAGT  65 2616 882721 N/A N/A 17224 17239 TTCCAGGTCATTTGAC  34 2617 882745 N/A N/A 17492 17507 CTAGATAGCATTCTTA  61 2618 882769 N/A N/A 17731 17746 CCACAGTTGAACCCAT  64 2619 882793 N/A N/A 17982 17997 TCAGAGCTAGAACATG  62 2620 882816 N/A N/A 18579 18594 GATTGATGTGATAGCT  44 2621 882840 N/A N/A 18960 18975 CTACTATTGTGGAAAA  97 2622 882863 N/A N/A 19230 19245 CTTAATTCTGGGTTAA  47 2623 882887 N/A N/A 19417 19432 ACATTACTGAGATATC  84 2624 882895 N/A N/A  5776  5791 CGAAAACAGCCGCCGG  85 2625 882901 N/A N/A  9358  9373 ACGCAGCCTCTAAGAA  75 2626

Example 7: Effect of Mixed MOE and cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured MM.1R cells at a density of 5,000 cells per well were transfected by free uptake with 1,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set hIRF4_LTS34726 (forward sequence GGCAAAGAAAGCTCATCACAG, designated herein as SEQ ID NO: 3389; reverse sequence GGATTGCTGATGTGTTCTGGTA designated herein as SEQ ID NO: 3390; probe sequence TAGCCCCTCAGGAAATGTCCACTG, designated herein as SEQ ID: 3391) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Table 37 are cEt and/or MOE containing gapmers. The modified oligonucleotides have a central gap segment comprising 2′-deoxynucleosides which is flanked by wing segments on the 5′ direction and the 3′ direction. At least one nucleoside in the 5′ wing segment and/or one nucleoside in the 3′ wing segment has a MOE and/or cEt sugar modification. The “Motif” column describes the sugar modifications of each oligonucleotide. “k” indicates a cEt sugar modification; “d” indicates deoxyribose; and “e” indicates a MOE modification. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Table 37 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 37 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 Compound Start Stop Start Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 47 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 15 2044 935895 4200 4215 22317 22332 TCTATAGTGTTCCAGG kk-d9-eeekk 50 1734 935896 3070 3085 21187 21202 GCTTGATAAAGGCTGA kk-d9-eeekk 72 1505 935897 3251 3266 21368 21383 CACTTTTAGAGAGGAG kk-d9-eeekk 67 2036 935898 4227 4242 22344 22359 GTTGTAAATGAGTCGG kk-d9-eeekk 18  559 935899 4591 4606 22708 22723 GAAGTTTACACTGGAT kk-d9-eeekk 72 1968 935900 N/A N/A  8464  8479 GCATTTTATCATCCGA kk-d9-eeekk 62 1605 935901 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kk-d9-eeekk 62 2627 935902 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kk-d9-eeekk 61  649 935903 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kk-d9-eeekk 68  548 935904 2914 2929 21031 21046 AGTATGAGAAACGGCC kk-d9-eeekk 84 1045 935905 4199 4214 22316 22331 CTATAGTGTTCCAGGA kk-d9-eeekk 66  552 935906 3069 3084 21186 21201 CTTGATAAAGGCTGAA kk-d9-eeekk 90 1427 935907 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kk-d9-eeekk 64 1960 935908 4226 4241 22343 22358 TTGTAAATGAGTCGGT kk-d9-eeekk 52  195 935909 4593 4608 22710 22725 CGGAAGTTTACACTGG kk-d9-eeekk 66 2119 935910 N/A N/A  8466  8481 AAGCATTTTATCATCC kk-d9-eeekk 79 2628 935911 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kk-d9-eeekk 44 2629 935912 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kk-d9-eeekk 67 1894 935913 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kk-d9-eeekk 55 1356 935914 2916 2931 21033 21048 GTAGTATGAGAAACGG kk-d9-eeekk 49  426 935915 4201 4216 22318 22333 CTCTATAGTGTTCCAG kk-d9-eeekk 52 1811 935916 3071 3086 21188 21203 AGCTTGATAAAGGCTG kk-d9-eeekk 97 1579 935917 3252 3267 21369 21384 ACACTTTTAGAGAGGA kk-d9-eeekk 79 2111 935918 4228 4243 22345 22360 AGTTGTAAATGAGTCG kk-d9-eeekk 34 2021 935919 4594 4609 22711 22726 ACGGAAGTTTACACTG kk-d9-eeekk 85 2195 935920 N/A N/A  8467  8482 GAAGCATTTTATCATC kk-d9-eeekk 91 2630 935921 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kk-d9-eeekk 19 2631 935922 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kk-d9-eeekk 96 1971 935923 4174 4189 22291 22306 TACTGCTTGAGGTTTT kk-d9-eeekk 90 1433 935924 2917 2932 21034 21049 TGTAGTATGAGAAACG kk-d9-eeekk 93 1197 935925 4202 4217 22319 22334 TCTCTATAGTGTTCCA kk-d9-eeekk 43 2632 935926 3072 3087 21189 21204 AAGCTTGATAAAGGCT kk-d9-eeekk 47 2633 935927 3253 3268 21370 21385 GACACTTTTAGAGAGG kk-d9-eeekk 75  451 935928 4229 4244 22346 22361 CAGTTGTAAATGAGTC kk-d9-eeekk 38  560 935929 4592 4607 22709 22724 GGAAGTTTACACTGGA kk-d9-ekeke 36 2044 935930 N/A N/A  8465  8480 AGCATTTTATCATCCG kk-d9-ekeke 54 1679 935931 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kk-d9-ekeke 67 1232 935932 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kk-d9-ekeke 72 1817 935933 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kk-d9-ekeke 51 1279 935934 2915 2930 21032 21047 TAGTATGAGAAACGGC kk-d9-ekeke 50 1121 935935 4200 4215 22317 22332 TCTATAGTGTTCCAGG kk-d9-ekeke 44 1734 935936 3070 3085 21187 21202 GCTTGATAAAGGCTGA kk-d9-ekeke 61 1505 935937 3251 3266 21368 21383 CACTTTTAGAGAGGAG kk-d9-ekeke 74 2036 935938 4227 4242 22344 22359 GTTGTAAATGAGTCGG kk-d9-ekeke 52  559 935939 4591 4606 22708 22723 GAAGTTTACACTGGAT kk-d9-ekeke 42 1968 935940 N/A N/A  8464  8479 GCATTTTATCATCCGA kk-d9-ekeke 56 1605 935941 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kk-d9-ekeke 42 2627 935942 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kk-d9-ekeke 55  649 935943 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kk-d9-ekeke 55  548 935944 2914 2929 21031 21046 AGTATGAGAAACGGCC kk-d9-ekeke 89 1045 935945 4199 4214 22316 22331 CTATAGTGTTCCAGGA kk-d9-ekeke 52  552 935946 3069 3084 21186 21201 CTTGATAAAGGCTGAA kk-d9-ekeke 81 1427 935947 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kk-d9-ekeke 48 1960 935948 4226 4241 22343 22358 TTGTAAATGAGTCGGT kk-d9-ekeke 43  195 935949 4593 4608 22710 22725 CGGAAGTTTACACTGG kk-d9-ekeke 67 2119 935950 N/A N/A  8466  8481 AAGCATTTTATCATCC kk-d9-ekeke 75 2628 935951 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kk-d9-ekeke 53 2629 935952 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kk-d9-ekeke 79 1894 935953 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kk-d9-ekeke 60 1356 935954 2916 2931 21033 21048 GTAGTATGAGAAACGG kk-d9-ekeke 65  426 935955 4201 4216 22318 22333 CTCTATAGTGTTCCAG kk-d9-ekeke 57 1811 935956 3071 3086 21188 21203 AGCTTGATAAAGGCTG kk-d9-ekeke 82 1579 935957 3252 3267 21369 21384 ACACTTTTAGAGAGGA kk-d9-ekeke 59 2111 935958 4228 4243 22345 22360 AGTTGTAAATGAGTCG kk-d9-ekeke 27 2021 935959 4594 4609 22711 22726 ACGGAAGTTTACACTG kk-d9-ekeke 92 2195 935960 N/A N/A  8467  8482 GAAGCATTTTATCATC kk-d9-ekeke 94 2630 935961 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kk-d9-ekeke 20 2631 935962 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kk-d9-ekeke 87 1971 935963 4174 4189 22291 22306 TACTGCTTGAGGTTTT kk-d9-ekeke 82 1433 935964 2917 2932 21034 21049 TGTAGTATGAGAAACG kk-d9-ekeke 87 1197 935965 4202 4217 22319 22334 TCTCTATAGTGTTCCA kk-d9-ekeke 61 2632 935966 3072 3087 21189 21204 AAGCTTGATAAAGGCT kk-d9-ekeke 62 2633 935967 3253 3268 21370 21385 GACACTTTTAGAGAGG kk-d9-ekeke 78  451 935968 4229 4244 22346 22361 CAGTTGTAAATGAGTC kk-d9-ekeke 28  560 935969 4592 4607 22709 22724 GGAAGTTTACACTGGA k-d9-kekeke 68 2044 935970 N/A N/A  8465  8480 AGCATTTTATCATCCG k-d9-kekeke 76 1679

Example 8: Effect of Mixed MOE and cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured MM.1R cells at a density of 5,000 cells per well were transfected by free uptake with 1,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 38 through 42 are cEt and/or MOE containing gapmers. The modified oligonucleotides have a central gap segment comprising 2′-deoxynucleosides which is flanked by wing segments on the 5′ direction and the 3′ direction. At least one nucleoside in the 5′ wing segment and/or one nucleoside in the 3′ wing segment has a MOE and/or cEt sugar modification. The “Motif” column describes the sugar modifications of each oligonucleotide. “k” indicates a cEt sugar modification; “d” indicates deoxyribose; and “e” indicates a MOE modification. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 38 through 42 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 38 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 Compound Start Stop Start Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk  37  195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk  53 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk  14 2044 935570 N/A N/A  8455  8470 CATCCGAAAAGACTGG kkk-d10-kkk  86 2634 935571 N/A N/A  8456  8471 TCATCCGAAAAGACTG kkk-d10-kkk  84 2635 935572 N/A N/A  8457  8472 ATCATCCGAAAAGACT kkk-d10-kkk  96 2636 935573 N/A N/A  8458  8473 TATCATCCGAAAAGAC kkk-d10-kkk  97 2637 935574 N/A N/A  8459  8474 TTATCATCCGAAAAGA kkk-d10-kkk  96 2638 935575 N/A N/A  8460  8475 TTTATCATCCGAAAAG kkk-d10-kkk  93 2639 935576 N/A N/A  8472  8487 CAGCCGAAGCATTTTA kkk-d10-kkk 100 2640 935577 N/A N/A  8473  8488 ACAGCCGAAGCATTTT kkk-d10-kkk  92 2641 935578 N/A N/A  8474  8489 GACAGCCGAAGCATTT kkk-d10-kkk  94 2642 935580 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kkk-d10-kkk  34 2627 935581 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kkk-d10-kkk  38 2629 935582 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kkk-d10-kkk  61 2631 935584 N/A N/A 11124 11139 CCACAAGCAGTGATGT kkk-d10-kkk  84 2643 935585 N/A N/A 11125 11140 ACCACAAGCAGTGATG kkk-d10-kkk  99 2644 935586 2905 2920 21022 21037 AACGGCCTGGAGGTGA kkk-d10-kkk  99 2645 935587 2906 2921 21023 21038 AAACGGCCTGGAGGTG kkk-d10-kkk  89 2646 935588 2907 2922 21024 21039 GAAACGGCCTGGAGGT kkk-d10-kkk  83 2647 935590 2920 2935 21037 21052 TCCTGTAGTATGAGAA kkk-d10-kkk  78 2648 935591 2922 2937 21039 21054 TATCCTGTAGTATGAG kkk-d10-kkk  58 2649 935593 3072 3087 21189 21204 AAGCTTGATAAAGGCT kkk-d10-kkk  97 2633 935597 3080 3095 21197 21212 TGCTCACTAAGCTTGA kkk-d10-kkk  88 2650 935603 4178 4193 22295 22310 TAATTACTGCTTGAGG kkk-d10-kkk  38 2651 935606 4194 4209 22311 22326 GTGTTCCAGGAGATAT kkk-d10-kkk  70 2652 935608 4196 4211 22313 22328 TAGTGTTCCAGGAGAT kkk-d10-kkk  47 2653 935611 4208 4223 22325 22340 CTTGGTTCTCTATAGT kkk-d10-kkk  73 2654 935614 4597 4612 22714 22729 TGAACGGAAGTTTACA kkk-d10-kkk  86 2655 935616 5190 5205 23307 23322 TACCTCAGTTTGTGAA kkk-d10-kkk  89 2656 935618 N/A N/A  8466  8481 AAGCATTTTATCATCC kkk-d10-kkk  75 2628 935619 N/A N/A  8467  8482 GAAGCATTTTATCATC kkk-d10-kkk  96 2630 935620 4202 4217 22319 22334 TCTCTATAGTGTTCCA kkk-d10-kkk  30 2632 935621 4592 4607 22709 22724 GGAAGTTTACACTGGA k-d10-kekek  67 2044 935622 N/A N/A  8465  8480 AGCATTTTATCATCCG k-d10-kekek  64 1679 935623 N/A N/A 11116 11131 AGTGATGTCAGGTTTT k-d10-kekek  75 1232 935624 5187 5202 23304 23319 CTCAGTTTGTGAAGCA k-d10-kekek  76 1817 935625 4172 4187 22289 22304 CTGCTTGAGGTTTTCC k-d10-kekek  92 1279 935626 2915 2930 21032 21047 TAGTATGAGAAACGGC k-d10-kekek  76 1121 935627 4200 4215 22317 22332 TCTATAGTGTTCCAGG k-d10-kekek  84 1734 935628 3070 3085 21187 21202 GCTTGATAAAGGCTGA k-d10-kekek  83 1505 935629 3251 3266 21368 21383 CACTTTTAGAGAGGAG k-d10-kekek  81 2036 935630 4591 4606 22708 22723 GAAGTTTACACTGGAT k-d10-kekek  73 1968 935631 N/A N/A  8464  8479 GCATTTTATCATCCGA k-d10-kekek  79 1605 935632 N/A N/A 11115 11130 GTGATGTCAGGTTTTC k-d10-kekek  78 2627 935633 5186 5201 23303 23318 TCAGTTTGTGAAGCAT k-d10-kekek  79  649 935634 4171 4186 22288 22303 TGCTTGAGGTTTTCCT k-d10-kekek  64  548 935635 2914 2929 21031 21046 AGTATGAGAAACGGCC k-d10-kekek  90 1045 935636 4199 4214 22316 22331 CTATAGTGTTCCAGGA k-d10-kekek  66  552 935637 3069 3084 21186 21201 CTTGATAAAGGCTGAA k-d10-kekek  83 1427 935638 3250 3265 21367 21382 ACTTTTAGAGAGGAGA k-d10-kekek  68 1960 935639 4593 4608 22710 22725 CGGAAGTTTACACTGG k-d10-kekek  90 2119 935640 N/A N/A  8466  8481 AAGCATTTTATCATCC k-d10-kekek  86 2628 935641 N/A N/A 11117 11132 CAGTGATGTCAGGTTT k-d10-kekek  54 2629 935642 5188 5203 23305 23320 CCTCAGTTTGTGAAGC k-d10-kekek  74 1894 935643 4173 4188 22290 22305 ACTGCTTGAGGTTTTC k-d10-kekek  78 1356 935644 2916 2931 21033 21048 GTAGTATGAGAAACGG k-d10-kekek  54  426 935645 4201 4216 22318 22333 CTCTATAGTGTTCCAG k-d10-kekek  76 1811 935646 3071 3086 21188 21203 AGCTTGATAAAGGCTG k-d10-kekek  99 1579 935647 3252 3267 21369 21384 ACACTTTTAGAGAGGA k-d10-kekek 106 2111 935648 4228 4243 22345 22360 AGTTGTAAATGAGTCG k-d10-kekek  56 2021 935649 4594 4609 22711 22726 ACGGAAGTTTACACTG k-d10-kekek  93 2195 935650 N/A N/A  8467 8482 GAAGCATTTTATCATC k-d10-kekek 112 2630 935651 N/A N/A 11118 11133 GCAGTGATGTCAGGTT k-d10-kekek  64 2631 935652 5189 5204 23306 23321 ACCTCAGTTTGTGAAG k-d10-kekek  82 1971 935653 4174 4189 22291 22306 TACTGCTTGAGGTTTT k-d10-kekek  76 1433 935654 2917 2932 21034 21049 TGTAGTATGAGAAACG k-d10-kekek  93 1197 935655 4202 4217 22319 22334 TCTCTATAGTGTTCCA k-d10-kekek  39 2632 935656 3072 3087 21189 21204 AAGCTTGATAAAGGCT k-d10-kekek  97 2633 935657 3253 3268 21370 21385 GACACTTTTAGAGAGG k-d10-kekek  77  451 935658 4229 4244 22346 22361 CAGTTGTAAATGAGTC k-d10-kekek  35  560 935659 4592 4607 22709 22724 GGAAGTTTACACTGGA kk-d9-kekek  53 2044 935660 N/A N/A   8465  8480 AGCATTTTATCATCCG kk-d9-kekek  61 1679 935661 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kk-d9-kekek  61 1232 935662 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kk-d9-kekek  78 1817 935663 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kk-d9-kekek  70 1279 935664 2915 2930 21032 21047 TAGTATGAGAAACGGC kk-d9-kekek  73 1121 935665 4200 4215 22317 22332 TCTATAGTGTTCCAGG kk-d9-kekek  72 1734 935666 3070 3085 21187 21202 GCTTGATAAAGGCTGA kk-d9-kekek  93 1505

TABLE 39 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 37 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 49 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 15 2044 935667 3251 3266 21368 21383 CACTTTTAGAGAGGAG kk-d9-kekek 83 2036 935668 4591 4606 22708 22723 GAAGTTTACACTGGAT kk-d9-kekek 45 1968 935669 N/A N/A 8464 8479 GCATTTTATCATCCGA kk-d9-kekek 52 1605 935670 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kk-d9-kekek 60 2627 935671 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kk-d9-kekek 42 649 935672 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kk-d9-kekek 64 548 935673 2914 2929 21031 21046 AGTATGAGAAACGGCC kk-d9-kekek 97 1045 935674 4199 4214 22316 22331 CTATAGTGTTCCAGGA kk-d9-kekek 57 552 935675 3069 3084 21186 21201 CTTGATAAAGGCTGAA kk-d9-kekek 88 1427 935676 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kk-d9-kekek 60 1960 935677 4593 4608 22710 22725 CGGAAGTTTACACTGG kk-d9-kekek 80 2119 935678 N/A N/A 8466 8481 AAGCATTTTATCATCC kk-d9-kekek 82 2628 935679 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kk-d9-kekek 36 2629 935680 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kk-d9-kekek 85 1894 935681 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kk-d9-kekek 76 1356 935682 2916 2931 21033 21048 GTAGTATGAGAAACGG kk-d9-kekek 50 426 935683 4201 4216 22318 22333 CTCTATAGTGTTCCAG kk-d9-kekek 78 1811 935684 3071 3086 21188 21203 AGCTTGATAAAGGCTG kk-d9-kekek 84 1579 935685 3252 3267 21369 21384 ACACTTTTAGAGAGGA kk-d9-kekek 73 2111 935686 4228 4243 22345 22360 AGTTGTAAATGAGTCG kk-d9-kekek 44 2021 935687 4594 4609 22711 22726 ACGGAAGTTTACACTG kk-d9-kekek 79 2195 935688 N/A N/A 8467 8482 GAAGCATTTTATCATC kk-d9-kekek 120 2630 935689 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kk-d9-kekek 31 2631 935690 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kk-d9-kekek 87 1971 935691 4174 4189 22291 22306 TACTGCTTGAGGTTTT kk-d9-kekek 78 1433 935692 2917 2932 21034 21049 TGTAGTATGAGAAACG kk-d9-kekek 88 1197 935693 4202 4217 22319 22334 TCTCTATAGTGTTCCA kk-d9-kekek 72 2632 935694 3072 3087 21189 21204 AAGCTTGATAAAGGCT kk-d9-kekek 119 2633 935695 3253 3268 21370 21385 GACACTTTTAGAGAGG kk-d9-kekek 83 451 935696 4229 4244 22346 22361 CAGTTGTAAATGAGTC kk-d9-kekek 19 560 935697 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d9-kkke 22 2044 935698 N/A N/A 8465 8480 AGCATTTTATCATCCG kkk-d9-kkke 34 1679 935699 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kkk-d9-kkke 35 1232 935700 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kkk-d9-kkke 31 1817 935701 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kkk-d9-kkke 35 1279 935702 2915 2930 21032 21047 TAGTATGAGAAACGGC kkk-d9-kkke 52 1121 935703 4200 4215 22317 22332 TCTATAGTGTTCCAGG kkk-d9-kkke 48 1734 935704 3070 3085 21187 21202 GCTTGATAAAGGCTGA kkk-d9-kkke 77 1505 935705 3251 3266 21368 21383 CACTTTTAGAGAGGAG kkk-d9-kkke 60 2036 935706 4591 4606 22708 22723 GAAGTTTACACTGGAT kkk-d9-kkke 55 1968 935707 N/A N/A 8464 8479 GCATTTTATCATCCGA kkk-d9-kkke 38 1605 935708 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kkk-d9-kkke 29 2627 935709 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kkk-d9-kkke 42 649 935710 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kkk-d9-kkke 54 548 935711 2914 2929 21031 21046 AGTATGAGAAACGGCC kkk-d9-kkke 93 1045 935712 4199 4214 22316 22331 CTATAGTGTTCCAGGA kkk-d9-kkke 69 552 935713 3069 3084 21186 21201 CTTGATAAAGGCTGAA kkk-d9-kkke 64 1427 935714 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kkk-d9-kkke 64 1960 935715 4593 4608 22710 22725 CGGAAGTTTACACTGG kkk-d9-kkke 75 2119 935716 N/A N/A 8466 8481 AAGCATTTTATCATCC kkk-d9-kkke 67 2628 935717 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kkk-d9-kkke 51 2629 935718 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kkk-d9-kkke 98 1894 935719 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kkk-d9-kkke 48 1356 935720 2916 2931 21033 21048 GTAGTATGAGAAACGG kkk-d9-kkke 60 426 935721 4201 4216 22318 22333 CTCTATAGTGTTCCAG kkk-d9-kkke 35 1811 935722 3071 3086 21188 21203 AGCTTGATAAAGGCTG kkk-d9-kkke 98 1579 935723 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d9-kkke 71 2111 935724 4228 4243 22345 22360 AGTTGTAAATGAGTCG kkk-d9-kkke 13 2021 935725 4594 4609 22711 22726 ACGGAAGTTTACACTG kkk-d9-kkke 71 2195 935726 N/A N/A 8467 8482 GAAGCATTTTATCATC kkk-d9-kkke 99 2630 935727 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kkk-d9-kkke 38 2631 935728 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kkk-d9-kkke 90 1971 935729 4174 4189 22291 22306 TACTGCTTGAGGTTTT kkk-d9-kkke 87 1433 935730 2917 2932 21034 21049 TGTAGTATGAGAAACG kkk-d9-kkke 67 1197 935731 4202 4217 22319 22334 TCTCTATAGTGTTCCA kkk-d9-kkke 44 2632 935732 3072 3087 21189 21204 AAGCTTGATAAAGGCT kkk-d9-kkke 95 2633 935733 3253 3268 21370 21385 GACACTTTTAGAGAGG kkk-d9-kkke 76 451 935734 4229 4244 22346 22361 CAGTTGTAAATGAGTC kkk-d9-kkke 32 560 935735 4592 4607 22709 22724 GGAAGTTTACACTGGA kk-d10-keke 54 2044 935736 N/A N/A 8465 8480 AGCATTTTATCATCCG kk-d10-keke 67 1679 935737 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kk-d10-keke 68 1232 935738 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kk-d10-keke 70 1817 935739 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kk-d10-keke 56 1279 935740 2915 2930 21032 21047 TAGTATGAGAAACGGC kk-d10-keke 54 1121 935741 4200 4215 22317 22332 TCTATAGTGTTCCAGG kk-d10-keke 34 1734 935742 3070 3085 21187 21202 GCTTGATAAAGGCTGA kk-d10-keke 69 1505

TABLE 40 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 37 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 53 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 20 2044 935743 3251 3266 21368 21383 CACTTTTAGAGAGGAG kk-d10-keke 73 2036 935744 4591 4606 22708 22723 GAAGTTTACACTGGAT kk-d10-keke 54 1968 935745 N/A N/A 8464 8479 GCATTTTATCATCCGA kk-d10-keke 61 1605 935746 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kk-d10-keke 82 2627 935747 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kk-d10-keke 67 649 935748 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kk-d10-keke 67 548 935749 2914 2929 21031 21046 AGTATGAGAAACGGCC kk-d10-keke 91 1045 935750 4199 4214 22316 22331 CTATAGTGTTCCAGGA kk-d10-keke 64 552 935751 3069 3084 21186 21201 CTTGATAAAGGCTGAA kk-d10-keke 96 1427 935752 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kk-d10-keke 77 1960 935753 4593 4608 22710 22725 CGGAAGTTTACACTGG kk-d10-keke 67 2119 935754 N/A N/A 8466 8481 AAGCATTTTATCATCC kk-d10-keke 80 2628 935755 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kk-d10-keke 59 2629 935756 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kk-d10-keke 84 1894 935757 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kk-d10-keke 79 1356 935758 2916 2931 21033 21048 GTAGTATGAGAAACGG kk-d10-keke 72 426 935759 4201 4216 22318 22333 CTCTATAGTGTTCCAG kk-d10-keke 73 1811 935760 3071 3086 21188 21203 AGCTTGATAAAGGCTG kk-d10-keke 99 1579 935761 3252 3267 21369 21384 ACACTTTTAGAGAGGA kk-d10-keke 86 2111 935762 4228 4243 22345 22360 AGTTGTAAATGAGTCG kk-d10-keke 41 2021 935763 4594 4609 22711 22726 ACGGAAGTTTACACTG kk-d10-keke 87 2195 935764 N/A N/A 8467 8482 GAAGCATTTTATCATC kk-d10-keke 107 2630 935765 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kk-d10-keke 33 2631 935766 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kk-d10-keke 112 1971 935767 4174 4189 22291 22306 TACTGCTTGAGGTTTT kk-d10-keke 89 1433 935768 2917 2932 21034 21049 TGTAGTATGAGAAACG kk-d10-keke 90 1197 935769 4202 4217 22319 22334 TCTCTATAGTGTTCCA kk-d10-keke 73 2632 935770 3072 3087 21189 21204 AAGCTTGATAAAGGCT kk-d10-keke 118 2633 935771 3253 3268 21370 21385 GACACTTTTAGAGAGG kk-d10-keke 91 451 935772 4229 4244 22346 22361 CAGTTGTAAATGAGTC kk-d10-keke 41 560 935773 4592 4607 22709 22724 GGAAGTTTACACTGGA kk-d9-kdkdk 76 2044 935774 N/A N/A 8465 8480 AGCATTTTATCATCCG kk-d9-kdkdk 89 1679 935775 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kk-d9-kdkdk 69 1232 935776 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kk-d9-kdkdk 101 1817 935777 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kk-d9-kdkdk 64 1279 935778 2915 2930 21032 21047 TAGTATGAGAAACGGC kk-d9-kdkdk 61 1121 935779 4200 4215 22317 22332 TCTATAGTGTTCCAGG kk-d9-kdkdk 50 1734 935780 3070 3085 21187 21202 GCTTGATAAAGGCTGA kk-d9-kdkdk 96 1505 935781 3251 3266 21368 21383 CACTTTTAGAGAGGAG kk-d9-kdkdk 84 2036 935782 4227 4242 22344 22359 GTTGTAAATGAGTCGG kk-d9-kdkdk 42 559 935783 4591 4606 22708 22723 GAAGTTTACACTGGAT kk-d9-kdkdk 77 1968 935784 N/A N/A 8464 8479 GCATTTTATCATCCGA kk-d9-kdkdk 84 1605 935785 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kk-d9-kdkdk 73 2627 935786 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kk-d9-kdkdk 75 649 935787 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kk-d9-kdkdk 55 548 935788 2914 2929 21031 21046 AGTATGAGAAACGGCC kk-d9-kdkdk 94 1045 935789 4199 4214 22316 22331 CTATAGTGTTCCAGGA kk-d9-kdkdk 51 552 935790 3069 3084 21186 21201 CTTGATAAAGGCTGAA kk-d9-kdkdk 99 1427 935791 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kk-d9-kdkdk 66 1960 935792 4226 4241 22343 22358 TTGTAAATGAGTCGGT kk-d9-kdkdk 57 195 935793 4593 4608 22710 22725 CGGAAGTTTACACTGG kk-d9-kdkdk 87 2119 935794 N/A N/A 8466 8481 AAGCATTTTATCATCC kk-d9-kdkdk 97 2628 935795 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kk-d9-kdkdk 53 2629 935796 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kk-d9-kdkdk 92 1894 935797 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kk-d9-kdkdk 81 1356 935798 2916 2931 21033 21048 GTAGTATGAGAAACGG kk-d9-kdkdk 68 426 935799 4201 4216 22318 22333 CTCTATAGTGTTCCAG kk-d9-kdkdk 69 1811 935800 3071 3086 21188 21203 AGCTTGATAAAGGCTG kk-d9-kdkdk 103 1579 935801 3252 3267 21369 21384 ACACTTTTAGAGAGGA kk-d9-kdkdk 97 2111 935802 4228 4243 22345 22360 AGTTGTAAATGAGTCG kk-d9-kdkdk 61 2021 935803 4594 4609 22711 22726 ACGGAAGTTTACACTG kk-d9-kdkdk 102 2195 935804 N/A N/A 8467 8482 GAAGCATTTTATCATC kk-d9-kdkdk 99 2630 935805 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kk-d9-kdkdk 47 2631 935806 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kk-d9-kdkdk 103 1971 935807 4174 4189 22291 22306 TACTGCTTGAGGTTTT kk-d9-kdkdk 85 1433 935808 2917 2932 21034 21049 TGTAGTATGAGAAACG kk-d9-kdkdk 106 1197 935809 4202 4217 22319 22334 TCTCTATAGTGTTCCA kk-d9-kdkdk 62 2632 935810 3072 3087 21189 21204 AAGCTTGATAAAGGCT kk-d9-kdkdk 102 2633 935811 3253 3268 21370 21385 GACACTTTTAGAGAGG kk-d9-kdkdk 86 451 935812 4229 4244 22346 22361 CAGTTGTAAATGAGTC kk-d9-kdkdk 54 560 935813 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d8-kekek 57 2044 935814 N/A N/A 8465 8480 AGCATTTTATCATCCG kkk-d8-kekek 78 1679 935815 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kkk-d8-kekek 63 1232 935816 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kkk-d8-kekek 63 1817 935817 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kkk-d8-kekek 74 1279 935818 2915 2930 21032 21047 TAGTATGAGAAACGGC kkk-d8-kekek 70 1121

TABLE 41 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 34 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 63 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 17 2044 935819 4200 4215 22317 22332 TCTATAGTGTTCCAGG kkk-d8-kekek 87 1734 935820 3070 3085 21187 21202 GCTTGATAAAGGCTGA kkk-d8-kekek 99 1505 935821 3251 3266 21368 21383 CACTTTTAGAGAGGAG kkk-d8-kekek 93 2036 935822 4591 4606 22708 22723 GAAGTTTACACTGGAT kkk-d8-kekek 48 1968 935823 N/A N/A 8464 8479 GCATTTTATCATCCGA kkk-d8-kekek 55 1605 935824 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kkk-d8-kekek 43 2627 935825 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kkk-d8-kekek 59 649 935826 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kkk-d8-kekek 73 548 935827 2914 2929 21031 21046 AGTATGAGAAACGGCC kkk-d8-kekek 90 1045 935828 4199 4214 22316 22331 CTATAGTGTTCCAGGA kkk-d8-kekek 77 552 935829 3069 3084 21186 21201 CTTGATAAAGGCTGAA kkk-d8-kekek 88 1427 935830 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kkk-d8-kekek 79 1960 935831 4593 4608 22710 22725 CGGAAGTTTACACTGG kkk-d8-kekek 82 2119 935832 N/A N/A 8466 8481 AAGCATTTTATCATCC kkk-d8-kekek 90 2628 935833 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kkk-d8-kekek 44 2629 935834 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kkk-d8-kekek 97 1894 935835 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kkk-d8-kekek 68 1356 935836 2916 2931 21033 21048 GTAGTATGAGAAACGG kkk-d8-kekek 57 426 935837 4201 4216 22318 22333 CTCTATAGTGTTCCAG kkk-d8-kekek 85 1811 935838 3071 3086 21188 21203 AGCTTGATAAAGGCTG kkk-d8-kekek 108 1579 935839 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d8-kekek 85 2111 935840 4228 4243 22345 22360 AGTTGTAAATGAGTCG kkk-d8-kekek 39 2021 935841 4594 4609 22711 22726 ACGGAAGTTTACACTG kkk-d8-kekek 86 2195 935842 N/A N/A 8467 8482 GAAGCATTTTATCATC kkk-d8-kekek 105 2630 935843 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kkk-d8-kekek 64 2631 935844 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kkk-d8-kekek 103 1971 935845 4174 4189 22291 22306 TACTGCTTGAGGTTTT kkk-d8-kekek 108 1433 935846 2917 2932 21034 21049 TGTAGTATGAGAAACG kkk-d8-kekek 77 1197 935847 4202 4217 22319 22334 TCTCTATAGTGTTCCA kkk-d8-kekek 92 2632 935848 3072 3087 21189 21204 AAGCTTGATAAAGGCT kkk-d8-kekek 104 2633 935849 3253 3268 21370 21385 GACACTTTTAGAGAGG kkk-d8-kekek 108 451 935850 4229 4244 22346 22361 CAGTTGTAAATGAGTC kkk-d8-kekek 24 560 935851 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d9-keke 22 2044 935852 N/A N/A 8465 8480 AGCATTTTATCATCCG kkk-d9-keke 58 1679 935853 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kkk-d9-keke 41 1232 935854 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kkk-d9-keke 36 1817 935855 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kkk-d9-keke 50 1279 935856 2915 2930 21032 21047 TAGTATGAGAAACGGC kkk-d9-keke 38 1121 935857 4200 4215 22317 22332 TCTATAGTGTTCCAGG kkk-d9-keke 32 1734 935858 3070 3085 21187 21202 GCTTGATAAAGGCTGA kkk-d9-keke 76 1505 935859 3251 3266 21368 21383 CACTTTTAGAGAGGAG kkk-d9-keke 43 2036 935860 4591 4606 22708 22723 GAAGTTTACACTGGAT kkk-d9-keke 51 1968 935861 N/A N/A 8464 8479 GCATTTTATCATCCGA kkk-d9-keke 68 1605 935862 N/A N/A 11115 11130 GTGATGTCAGGTTTTC kkk-d9-keke 66 2627 935863 5186 5201 23303 23318 TCAGTTTGTGAAGCAT kkk-d9-keke 47 649 935864 4171 4186 22288 22303 TGCTTGAGGTTTTCCT kkk-d9-keke 65 548 935865 2914 2929 21031 21046 AGTATGAGAAACGGCC kkk-d9-keke 84 1045 935866 4199 4214 22316 22331 CTATAGTGTTCCAGGA kkk-d9-keke 78 552 935867 3069 3084 21186 21201 CTTGATAAAGGCTGAA kkk-d9-keke 76 1427 935868 3250 3265 21367 21382 ACTTTTAGAGAGGAGA kkk-d9-keke 59 1960 935869 4593 4608 22710 22725 CGGAAGTTTACACTGG kkk-d9-keke 58 2119 935870 N/A N/A 8466 8481 AAGCATTTTATCATCC kkk-d9-keke 79 2628 935871 N/A N/A 11117 11132 CAGTGATGTCAGGTTT kkk-d9-keke 66 2629 935872 5188 5203 23305 23320 CCTCAGTTTGTGAAGC kkk-d9-keke 92 1894 935873 4173 4188 22290 22305 ACTGCTTGAGGTTTTC kkk-d9-keke 62 1356 935874 2916 2931 21033 21048 GTAGTATGAGAAACGG kkk-d9-keke 85 426 935875 4201 4216 22318 22333 CTCTATAGTGTTCCAG kkk-d9-keke 51 1811 935876 3071 3086 21188 21203 AGCTTGATAAAGGCTG kkk-d9-keke 108 1579 935877 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d9-keke 63 2111 935878 4228 4243 22345 22360 AGTTGTAAATGAGTCG kkk-d9-keke 23 2021 935879 4594 4609 22711 22726 ACGGAAGTTTACACTG kkk-d9-keke 87 2195 935880 N/A N/A 8467 8482 GAAGCATTTTATCATC kkk-d9-keke 92 2630 935881 N/A N/A 11118 11133 GCAGTGATGTCAGGTT kkk-d9-keke 44 2631 935882 5189 5204 23306 23321 ACCTCAGTTTGTGAAG kkk-d9-keke 92 1971 935883 4174 4189 22291 22306 TACTGCTTGAGGTTTT kkk-d9-keke 93 1433 935884 2917 2932 21034 21049 TGTAGTATGAGAAACG kkk-d9-keke 71 1197 935885 4202 4217 22319 22334 TCTCTATAGTGTTCCA kkk-d9-keke 67 2632 935886 3072 3087 21189 21204 AAGCTTGATAAAGGCT kkk-d9-keke 118 2633 935887 3253 3268 21370 21385 GACACTTTTAGAGAGG kkk-d9-keke 76 451 935888 4229 4244 22346 22361 CAGTTGTAAATGAGTC kkk-d9-keke 42 560 935889 4592 4607 22709 22724 GGAAGTTTACACTGGA kk-d9-eeekk 53 2044 935890 N/A N/A 8465 8480 AGCATTTTATCATCCG kk-d9-eeekk 72 1679 935891 N/A N/A 11116 11131 AGTGATGTCAGGTTTT kk-d9-eeekk 75 1232 935892 5187 5202 23304 23319 CTCAGTTTGTGAAGCA kk-d9-eeekk 82 1817 935893 4172 4187 22289 22304 CTGCTTGAGGTTTTCC kk-d9-eeekk 69 1279 935894 2915 2930 21032 21047 TAGTATGAGAAACGGC kk-d9-eeekk 63 1121

TABLE 42 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID: IRF4 Compound 1 Start 1 Stop 2 Start 2 Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 31 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 54 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 15 2044 935971 N/A N/A 11116 11131 AGTGATGTCAGGTTTT k-d9-kekeke 60 1232 935972 5187 5202 23304 23319 CTCAGTTTGTGAAGCA k-d9-kekeke 81 1817 935973 4172 4187 22289 22304 CTGCTTGAGGTTTTCC k-d9-kekeke 80 1279 935974 2915 2930 21032 21047 TAGTATGAGAAACGGC k-d9-kekeke 68 1121 935975 4200 4215 22317 22332 TCTATAGTGTTCCAGG k-d9-kekeke 88 1734 935976 3070 3085 21187 21202 GCTTGATAAAGGCTGA k-d9-kekeke 101 1505 935977 3251 3266 21368 21383 CACTTTTAGAGAGGAG k-d9-kekeke 91 2036 935978 4591 4606 22708 22723 GAAGTTTACACTGGAT k-d9-kekeke 89 1968 935979 N/A N/A 8464 8479 GCATTTTATCATCCGA k-d9-kekeke 77 1605 935980 N/A N/A 11115 11130 GTGATGTCAGGTTTTC k-d9-kekeke 70 2627 935981 5186 5201 23303 23318 TCAGTTTGTGAAGCAT k-d9-kekeke 91 649 935982 4171 4186 22288 22303 TGCTTGAGGTTTTCCT k-d9-kekeke 91 548 935983 2914 2929 21031 21046 AGTATGAGAAACGGCC k-d9-kekeke 96 1045 935984 4199 4214 22316 22331 CTATAGTGTTCCAGGA k-d9-kekeke 88 552 935985 3069 3084 21186 21201 CTTGATAAAGGCTGAA k-d9-kekeke 95 1427 935986 3250 3265 21367 21382 ACTTTTAGAGAGGAGA k-d9-kekeke 83 1960 935987 4593 4608 22710 22725 CGGAAGTTTACACTGG k-d9-kekeke 83 2119 935988 N/A N/A 8466 8481 AAGCATTTTATCATCC k-d9-kekeke 94 2628 935989 N/A N/A 11117 11132 CAGTGATGTCAGGTTT k-d9-kekeke 52 2629 935990 5188 5203 23305 23320 CCTCAGTTTGTGAAGC k-d9-kekeke 89 1894 935991 4173 4188 22290 22305 ACTGCTTGAGGTTTTC k-d9-kekeke 79 1356 935992 2916 2931 21033 21048 GTAGTATGAGAAACGG k-d9-kekeke 72 426 935993 4201 4216 22318 22333 CTCTATAGTGTTCCAG k-d9-kekeke 89 1811 935994 3071 3086 21188 21203 AGCTTGATAAAGGCTG k-d9-kekeke 98 1579 935995 3252 3267 21369 21384 ACACTTTTAGAGAGGA k-d9-kekeke 94 2111 935996 4228 4243 22345 22360 AGTTGTAAATGAGTCG k-d9-kekeke 53 2021 935997 4594 4609 22711 22726 ACGGAAGTTTACACTG k-d9-kekeke 98 2195 935998 N/A N/A 8467 8482 GAAGCATTTTATCATC k-d9-kekeke 96 2630 935999 N/A N/A 11118 11133 GCAGTGATGTCAGGTT k-d9-kekeke 67 2631 936000 5189 5204 23306 23321 ACCTCAGTTTGTGAAG k-d9-kekeke 84 1971 936001 4174 4189 22291 22306 TACTGCTTGAGGTTTT k-d9-kekeke 86 1433 936002 2917 2932 21034 21049 TGTAGTATGAGAAACG k-d9-kekeke 86 1197 936003 4202 4217 22319 22334 TCTCTATAGTGTTCCA k-d9-kekeke 82 2632 936004 3072 3087 21189 21204 AAGCTTGATAAAGGCT k-d9-kekeke 98 2633 936005 3253 3268 21370 21385 GACACTTTTAGAGAGG k-d9-kekeke 91 451 936006 4229 4244 22346 22361 CAGTTGTAAATGAGTC k-d9-kekeke 38 560 936007 4592 4607 22709 22724 GGAAGTTTACACTGGA ekk-d9-kkee 45 2044 936008 N/A N/A 8465 8480 AGCATTTTATCATCCG ekk-d9-kkee 57 1679 936009 N/A N/A 11116 11131 AGTGATGTCAGGTTTT ekk-d9-kkee 61 1232 936010 5187 5202 23304 23319 CTCAGTTTGTGAAGCA ekk-d9-kkee 55 1817 936011 4172 4187 22289 22304 CTGCTTGAGGTTTTCC ekk-d9-kkee 48 1279 936012 2915 2930 21032 21047 TAGTATGAGAAACGGC ekk-d9-kkee 62 1121 936013 4200 4215 22317 22332 TCTATAGTGTTCCAGG ekk-d9-kkee 42 1734 936014 3070 3085 21187 21202 GCTTGATAAAGGCTGA ekk-d9-kkee 65 1505 936015 3251 3266 21368 21383 CACTTTTAGAGAGGAG ekk-d9-kkee 59 2036 936016 4227 4242 22344 22359 GTTGTAAATGAGTCGG ekk-d9-kkee 31 559 936017 4591 4606 22708 22723 GAAGTTTACACTGGAT ekk-d9-kkee 52 1968 936018 N/A N/A 8464 8479 GCATTTTATCATCCGA ekk-d9-kkee 39 1605 936019 N/A N/A 11115 11130 GTGATGTCAGGTTTTC ekk-d9-kkee 52 2627 936020 5186 5201 23303 23318 TCAGTTTGTGAAGCAT ekk-d9-kkee 53 649 936021 4171 4186 22288 22303 TGCTTGAGGTTTTCCT ekk-d9-kkee 65 548 936022 2914 2929 21031 21046 AGTATGAGAAACGGCC ekk-d9-kkee 88 1045 936023 4199 4214 22316 22331 CTATAGTGTTCCAGGA ekk-d9-kkee 92 552 936024 3069 3084 21186 21201 CTTGATAAAGGCTGAA ekk-d9-kkee 83 1427 936025 3250 3265 21367 21382 ACTTTTAGAGAGGAGA ekk-d9-kkee 72 1960 936026 4226 4241 22343 22358 TTGTAAATGAGTCGGT ekk-d9-kkee 49 195 936027 4593 4608 22710 22725 CGGAAGTTTACACTGG ekk-d9-kkee 76 2119 936028 N/A N/A 8466 8481 AAGCATTTTATCATCC ekk-d9-kkee 86 2628 936029 N/A N/A 11117 11132 CAGTGATGTCAGGTTT ekk-d9-kkee 80 2629 936030 5188 5203 23305 23320 CCTCAGTTTGTGAAGC ekk-d9-kkee 89 1894 936031 4173 4188 22290 22305 ACTGCTTGAGGTTTTC ekk-d9-kkee 54 1356 936032 2916 2931 21033 21048 GTAGTATGAGAAACGG ekk-d9-kkee 71 426 936033 4201 4216 22318 22333 CTCTATAGTGTTCCAG ekk-d9-kkee 41 1811 936034 3071 3086 21188 21203 AGCTTGATAAAGGCTG ekk-d9-kkee 99 1579 936035 3252 3267 21369 21384 ACACTTTTAGAGAGGA ekk-d9-kkee 71 2111 936036 4228 4243 22345 22360 AGTTGTAAATGAGTCG ekk-d9-kkee 51 2021 936037 4594 4609 22711 22726 ACGGAAGTTTACACTG ekk-d9-kkee 83 2195 936038 N/A N/A 8467 8482 GAAGCATTTTATCATC ekk-d9-kkee 99 2630 936039 N/A N/A 11118 11133 GCAGTGATGTCAGGTT ekk-d9-kkee 44 2631 936040 5189 5204 23306 23321 ACCTCAGTTTGTGAAG ekk-d9-kkee 98 1971 936041 4174 4189 22291 22306 TACTGCTTGAGGTTTT ekk-d9-kkee 94 1433 936042 2917 2932 21034 21049 TGTAGTATGAGAAACG ekk-d9-kkee 94 1197 936043 4202 4217 22319 22334 TCTCTATAGTGTTCCA ekk-d9-kkee 61 2632 936044 3072 3087 21189 21204 AAGCTTGATAAAGGCT ekk-d9-kkee 101 2633 936045 3253 3268 21370 21385 GACACTTTTAGAGAGG ekk-d9-kkee 76 451 936046 4229 4244 22346 22361 CAGTTGTAAATGAGTC ekk-d9-kkee 45 560

Example 9: Effect of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured MM.1R cells at a density of 5,000 cells per well were transfected by free uptake with 1,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the table below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 43 through 52 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 43 through 52 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 43 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 37 195 969844 N/A N/A 3801 3816 CCTTACCTCGCCCTGG 131 2657 969854 N/A N/A 4265 4280 CGCCAGCGGGTGAGCA 81 2658 969864 N/A N/A 4371 4386 GGCGACGACAGCTGCG 115 2659 969874 N/A N/A 4518 4533 AGCTAGCGCGCACTAA 108 2660 969884 N/A N/A 4836 4851 TCCTGTAACGCACCCG 113 2661 969894 N/A N/A 5487 5502 CCTGATGCCTCCGCCG 94 2662 969904 N/A N/A 5667 5682 CGAGAACGCACGGACG 111 2663 969914 N/A N/A 5761 5776 GCACAGGCGCGGACGC 106 2664 969924 N/A N/A 6002 6017 GTGCACTCGCGCAAAG 105 2665 969934 N/A N/A 6266 6281 TGCGGAGGTTCCTTGA 88 2666 969944 N/A N/A 6317 6332 CTGCCAAGTTGAAGAC 67 2667 969954 N/A N/A 6407 6422 GTCCTTCAGATTTACA 117 2668 969964 N/A N/A 6771 6786 ATACATGTCTGGTTTA 96 2669 969974 511 526 6990 7005 TTTTGGCTCCCTCAGG 103 2670 969984 N/A N/A 7169 7184 TTTCAACTTGTGACCC 99 2671 969994 N/A N/A 7222 7237 ATACATTGAGGCATAC 98 2672 970004 N/A N/A 7577 7592 GACATAAAGGACCCCG 92 2673 970014 N/A N/A 7639 7654 GGTCTGAGTTGTACAG 80 2674 970024 N/A N/A 8104 8119 CACCAATGGCAGCACC 84 2675 970034 N/A N/A 8221 8236 CATGATAAGGCACTAC 106 2676 970044 N/A N/A 8384 8399 TGGAGATACTTGTACT 58 2677 970054 N/A N/A 8489 8504 ATCCCTTATTAGACTG 107 2678 970064 645 660 9135 9150 CCAGCTTCGGTCGAGG 105 2679 970074 701 716 9191 9206 GTCATGGGACATTGGT 74 2680 970084 N/A N/A 9431 9446 CTGAGAGTAAACTTGG 101 2681 970094 N/A N/A 9582 9597 GCTCAATAATCTCCCA 100 2682 970104 N/A N/A 9679 9694 GTGTTTGCCATGGTAT 43 2683 970114 N/A N/A 9842 9857 CGCATTGCTAGATTCT 63 2684 970124 N/A N/A 9987 10002 GGATAACCTGAACATG 91 2685 970134 N/A N/A 10120 10135 CCATATTGGAAACCAG 90 2686 970144 N/A N/A 10178 10193 CAGTAAACGCAAGTCT 114 2687 970154 N/A N/A 10262 10277 AGACAGGTCTCTACCT 112 2688 970164 N/A N/A 10449 10464 TGCCAAAGAGCCCAAT 119 2689 970174 N/A N/A 10675 10690 AACTAGCAGGGCACGC 98 2690 970184 811 826 10876 10891 GGACTCCGGGAGCCTG 108 2691 970194 N/A N/A 11103 11118 TTTCTAATGGTGCTCC 96 2692 970203 N/A N/A 11365 11380 AACTAATGTCCCCAGG 112 2693 970213 N/A N/A 11452 11467 TTGTTTGCAAGCTATA 64 2694 970223 N/A N/A 11540 11555 TTCTTTATAGTAGGTA 85 2695 970233 N/A N/A 11664 11679 GAATTCCAAACCTTAA 96 2696 970243 N/A N/A 11946 11961 ACACAAGTCTTAGGTG 253 2697 970253 N/A N/A 12007 12022 TCAGAAATCACGAGGT 57 2698 970263 N/A N/A 12213 12228 TAATCTGTATCATGCA 78 2699 970273 N/A N/A 12293 12308 AGCTGCCACTGGTAAC 100 2700 970283 N/A N/A 12676 12691 GGCCTTAATGGTGATT 116 2701 970293 N/A N/A 12929 12944 GAAATGAACCCTAAGT 105 2702 970303 N/A N/A 13129 13144 ACCGACTCTTTTTTTA 102 2703 970313 N/A N/A 13400 13415 CAGAGCTCCGGAGTCA 106 2704 970323 N/A N/A 14015 14030 GCGACTGCTGAAAACC 104 2705 970333 N/A N/A 14206 14221 AGTACAGTCCACTCCA 93 2706 970343 N/A N/A 14248 14263 CCAACTTATAGCACTC 86 2707 970353 N/A N/A 14673 14688 ACAAAAACTTGGGTCA 113 2708 970363 N/A N/A 15188 15203 GATGGGACCGCCCTGG 113 2709 970373 N/A N/A 15597 15612 CTAGTCGCGCAAGTCT 104 2710 970383 N/A N/A 15852 15867 CAGAATGGCGAGTTGG 69 2711 970393 N/A N/A 15918 15933 CTTAGTCAGAATCTGT 99 2712 970403 N/A N/A 16174 16189 TAAAATGTCACGCCCG 100 2713 970413 N/A N/A 16468 16483 CACCAGCCATCGGCAG 119 2714 970423 N/A N/A 16699 16714 AGTGAAGTCGGGAGAT 92 2715 970433 N/A N/A 16869 16884 GGCTCTTGATGTGAAC 112 2716 970443 N/A N/A 16961 16976 ATCACCGAACACACCA 112 2717

TABLE 44 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 26 195 935583 N/A N/A 11119 11134 AGCAGTGATGTCAGGT 16 2718 969845 N/A N/A 4005 4020 GCCAAGGAGGCGGGCC 102 2719 969855 N/A N/A 4294 4309 ACCCAAATGTGGAGCT 97 2720 969865 N/A N/A 4377 4392 GGAGAGGGCGACGACA 101 2721 969875 N/A N/A 4541 4556 GAATAGGACCCCTATC 103 2722 969885 N/A N/A 4876 4891 TCCGAGCTCGGCCCCC 110 2723 969895 N/A N/A 5506 5521 TGCGGCTCCGGCGACG 120 2724 969905 N/A N/A 5670 5685 AAACGAGAACGCACGG 91 2725 969915 N/A N/A 5766 5781 CGCCGGCACAGGCGCG 86 2726 969925 N/A N/A 6006 6021 GTGTGTGCACTCGCGC 81 2727 969935 N/A N/A 6269 6284 AGATGCGGAGGTTCCT 68 2728 969945 N/A N/A 6324 6339 CCTATCACTGCCAAGT 103 2729 969955 N/A N/A 6412 6427 CATAGGTCCTTCAGAT 105 2730 969965 414 429 6893 6908 CAAAGCGCACCGCAGG 104 2731 969975 N/A N/A 6999 7014 CCCTACCTTTTTTGGC 107 2732 969985 N/A N/A 7186 7201 GCTGAACCCCACAGGA 100 2733 969995 N/A N/A 7223 7238 TATACATTGAGGCATA 93 2734 970005 N/A N/A 7579 7594 GTGACATAAAGGACCC 90 2735 970015 N/A N/A 7641 7656 AAGGTCTGAGTTGTAC 80 2736 970025 N/A N/A 8130 8145 CCCGACCCTCCCCAAC 135 2737 970035 N/A N/A 8223 8238 CACATGATAAGGCACT 79 2738 970045 N/A N/A 8387 8402 CAATGGAGATACTTGT 116 2739 970055 N/A N/A 8493 8508 GGGAATCCCTTATTAG 100 2740 970065 647 662 9137 9152 CTCCAGCTTCGGTCGA 91 2741 970075 N/A N/A 9280 9295 AGCAATTAGCTCTTCT 89 2742 970085 N/A N/A 9435 9450 GATCCTGAGAGTAAAC 94 2743 970095 N/A N/A 9583 9598 AGCTCAATAATCTCCC 94 2744 970105 N/A N/A 9740 9755 CCACAATCAGCAAGTC 100 2745 970115 N/A N/A 9844 9859 ACCGCATTGCTAGATT 73 2746 970125 N/A N/A 9995 10010 GCAGCCAAGGATAACC 94 2747 970135 N/A N/A 10127 10142 TCCACTACCATATTGG 129 2748 970145 N/A N/A 10180 10195 AGCAGTAAACGCAAGT 73 2749 970155 N/A N/A 10268 10283 GCTTCAAGACAGGTCT 71 2750 970165 N/A N/A 10477 10492 GCTGAGAGTTCAGGTC 93 2751 970175 N/A N/A 10678 10693 AGCAACTAGCAGGGCA 102 2752 970185 N/A N/A 11001 11016 TCGAATCTGCCCAAAG 77 2753 970204 N/A N/A 11369 11384 CCAGAACTAATGTCCC 94 2754 970214 N/A N/A 11455 11470 TATTTGTTTGCAAGCT 70 2755 970224 N/A N/A 11549 11564 AGAGGTGCCTTCTTTA 82 2756 970234 N/A N/A 11749 11764 CCACAACTCTCGCCTC 109 2757 970244 N/A N/A 11948 11963 CCACACAAGTCTTAGG 85 2758 970254 N/A N/A 12048 12063 CGAGGTGATTCTCGGG 85 2759 970264 N/A N/A 12222 12237 GCTGATAATTAATCTG 109 2760 970274 N/A N/A 12373 12388 CGCCCATGAGTTGAAA 87 2761 970284 N/A N/A 12692 12707 AAAGGGTAAGCACTGA 81 2762 970294 N/A N/A 12991 13006 ATTTAAGTCATGTGTC 83 2763 970304 N/A N/A 13132 13147 GCCACCGACTCTTTTT 105 2764 970314 N/A N/A 13416 13431 CGGCAGTCTGCAAACA 76 2765 970324 N/A N/A 14016 14031 GGCGACTGCTGAAAAC 110 2766 970334 N/A N/A 14207 14222 CAGTACAGTCCACTCC 95 2767 970344 N/A N/A 14249 14264 GCCAACTTATAGCACT 62 2768 970354 N/A N/A 14686 14701 TCTGGATGAGCTTACA 86 2769 970364 N/A N/A 15375 15390 ATCCACTGGCACCAAG 95 2770 970374 N/A N/A 15599 15614 TTCTAGTCGCGCAAGT 107 2771 970384 N/A N/A 15866 15881 CTACACAGGCTAATCA 96 2772 970394 N/A N/A 15920 15935 GACTTAGTCAGAATCT 90 2773 970404 N/A N/A 16176 16191 AATAAAATGTCACGCC 80 2774 970414 N/A N/A 16610 16625 CAGAATGTTTCGACAT 86 2775 970424 N/A N/A 16703 16718 CCACAGTGAAGTCGGG 94 2776 970434 N/A N/A 16885 16900 TTACTCCGCTGAGTGG 104 2777 970444 N/A N/A 16964 16979 CTCATCACCGAACACA 92 2778

TABLE 45 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 33 195 969846 N/A N/A 4037 4052 GCGCGGAGGGCAGGCG 132 2779 969856 N/A N/A 4298 4313 AGCGACCCAAATGTGG 102 2780 969866 N/A N/A 4407 4422 GCCCGGGAGAGCGGAG 132 2781 969876 N/A N/A 4637 4652 CGCGGAGGACCTCGCC 107 2782 969886 N/A N/A 4896 4911 GCGGGCACAGCCGTCC 121 2783 969896 N/A N/A 5530 5545 AGCCGAGGCCTCCTTT 128 2784 969906 N/A N/A 5673 5688 TGGAAACGAGAACGCA 107 2785 969916 N/A N/A 5774 5789 AAAACAGCCGCCGGCA 109 2786 969926 N/A N/A 6022 6037 CGTAACAACGACACAC 135 2787 969936 N/A N/A 6272 6287 GTGAGATGCGGAGGTT 61 2788 969946 N/A N/A 6360 6375 AGCTATGCTCTAGGAA 89 2789 969956 N/A N/A 6414 6429 CGCATAGGTCCTTCAG 94 2790 969966 429 444 6908 6923 GTCATTGCTCTTGTTC 82 2791 969976 N/A N/A 7004 7019 AGAGCCCCTACCTTTT 107 2792 969986 N/A N/A 7188 7203 ATGCTGAACCCCACAG 84 2793 969996 N/A N/A 7224 7239 ATATACATTGAGGCAT 131 2794 970006 N/A N/A 7592 7607 GCGAATGTGCCTTGTG 88 2795 970016 N/A N/A 7644 7659 TACAAGGTCTGAGTTG 106 2796 970026 N/A N/A 8132 8147 CGCCCGACCCTCCCCA 124 2797 970036 N/A N/A 8224 8239 TCACATGATAAGGCAC 81 2798 970046 N/A N/A 8390 8405 GGACAATGGAGATACT 97 2799 970056 N/A N/A 8496 8511 TCAGGGAATCCCTTAT 96 2800 970066 650 665 9140 9155 TCCCTCCAGCTTCGGT 121 2801 970076 N/A N/A 9284 9299 CATTAGCAATTAGCTC 137 2802 970086 N/A N/A 9437 9452 TGGATCCTGAGAGTAA 100 2803 970096 N/A N/A 9587 9602 TACCAGCTCAATAATC 127 2804 970106 N/A N/A 9744 9759 TAATCCACAATCAGCA 127 2805 970116 N/A N/A 9847 9862 GTTACCGCATTGCTAG 84 2806 970126 N/A N/A 10002 10017 ACTACCTGCAGCCAAG 96 2807 970136 N/A N/A 10130 10145 TCCTCCACTACCATAT 113 2808 970146 N/A N/A 10184 10199 CCAGAGCAGTAAACGC 89 2809 970156 N/A N/A 10273 10288 CCGATGCTTCAAGACA 87 2810 970166 N/A N/A 10489 10504 GTTCACAACAGAGCTG 116 2811 970176 N/A N/A 10711 10726 CGCCCAATCACCTTCC 119 2812 970186 N/A N/A 11005 11020 CCCATCGAATCTGCCC 124 2813 970195 N/A N/A 11127 11142 GAACCACAAGCAGTGA 132 2814 970205 N/A N/A 11371 11386 GACCAGAACTAATGTC 127 2815 970215 N/A N/A 11468 11483 CAGATTGAATCCATAT 80 2816 970225 N/A N/A 11553 11568 GCCTAGAGGTGCCTTC 87 2817 970235 N/A N/A 11797 11812 AAAGAGCTGGTAGGTC 135 2818 970245 N/A N/A 11960 11975 CTCTTCAGGCACCCAC 87 2819 970255 N/A N/A 12052 12067 CATTCGAGGTGATTCT 89 2820 970265 N/A N/A 12224 12239 TGGCTGATAATTAATC 105 2821 970275 N/A N/A 12383 12398 TTTAAAATATCGCCCA 101 2822 970285 N/A N/A 12694 12709 TTAAAGGGTAAGCACT 118 2823 970295 N/A N/A 13039 13054 ACTTGCTAAGTCTTAT 87 2824 970305 N/A N/A 13197 13212 TGGAGAAGTCCCGTGG 109 2825 970315 N/A N/A 13769 13784 CCTTACCTGACAAGAA 111 2826 970325 N/A N/A 14019 14034 AGAGGCGACTGCTGAA 119 2827 970335 N/A N/A 14208 14223 CCAGTACAGTCCACTC 106 2828 970345 N/A N/A 14332 14347 CGCTGAATTGTCATGA 101 2829 970355 N/A N/A 14688 14703 AATCTGGATGAGCTTA 96 2830 970365 N/A N/A 15410 15425 TCATTAGAAAGCCCTC 119 2831 970375 N/A N/A 15602 15617 AAGTTCTAGTCGCGCA 89 2832 970385 N/A N/A 15868 15883 ACCTACACAGGCTAAT 104 2833 970395 N/A N/A 15986 16001 GCTAACTTACAGGACT 108 2834 970405 N/A N/A 16252 16267 AGACAAGTGCCCATCC 99 2835 970415 N/A N/A 16611 16626 TCAGAATGTTTCGACA 94 2836 970425 N/A N/A 16712 16727 GGTAGTAGACCACAGT 77 2837 970435 N/A N/A 16890 16905 CACTCTTACTCCGCTG 112 2838 970445 N/A N/A 16966 16981 CCCTCATCACCGAACA 125 2839

TABLE 46 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 33 195 969847 N/A N/A 4044 4059 CGCAGGAGCGCGGAGG 128 2840 969857 N/A N/A 4302 4317 TCGGAGCGACCCAAAT 88 2841 969867 N/A N/A 4444 4459 ATTCCGCGCGCAGAGC 121 2842 969877 N/A N/A 4643 4658 TCCACGCGCGGAGGAC 143 2843 969887 N/A N/A 4928 4943 ACCTTCGCGGCCGGCC 142 2844 969897 N/A N/A 5570 5585 CGCCCAGGACCCGGCT 101 2845 969907 N/A N/A 5705 5720 CGGGAGCCCGGAGGAA 103 2846 969917 N/A N/A 5782 5797 GAGAGACGAAAACAGC 95 2847 969927 N/A N/A 6112 6127 GAGGAAGTCCCCTTCC 96 2848 969937 N/A N/A 6274 6289 GAGTGAGATGCGGAGG 45 2849 969947 N/A N/A 6364 6379 CCCCAGCTATGCTCTA 130 2850 969957 N/A N/A 6416 6431 GGCGCATAGGTCCTTC 107 2851 969967 446 461 6925 6940 TCAACCAGTTCCTCAA 123 2852 969977 N/A N/A 7008 7023 CAGGAGAGCCCCTACC 108 2853 969987 N/A N/A 7191 7206 CCTATGCTGAACCCCA 112 2854 969997 N/A N/A 7225 7240 CATATACATTGAGGCA 80 2855 970007 N/A N/A 7598 7613 TGGCATGCGAATGTGC 121 2856 970017 N/A N/A 7648 7663 TTTCTACAAGGTCTGA 97 2857 970027 N/A N/A 8135 8150 ACACGCCCGACCCTCC 96 2858 970037 N/A N/A 8231 8246 TGTGGTTTCACATGAT 79 2859 970047 N/A N/A 8403 8418 GGAGGATCATAAAGGA 68 2860 970057 N/A N/A 8520 8535 GTGCTCTTACAGCCTC 115 2861 970067 655 670 9145 9160 CGTAGTCCCTCCAGCT 73 2862 970077 N/A N/A 9289 9304 GGCCACATTAGCAATT 100 2863 970087 N/A N/A 9440 9455 GCATGGATCCTGAGAG 89 2864 970097 N/A N/A 9600 9615 TTCGAGAGAAATATAC 78 2865 970107 N/A N/A 9748 9763 CATCTAATCCACAATC 119 2866 970117 N/A N/A 9849 9864 GAGTTACCGCATTGCT 53 2867 970127 N/A N/A 10004 10019 TCACTACCTGCAGCCA 94 2868 970137 N/A N/A 10133 10148 AATTCCTCCACTACCA 141 2869 970147 N/A N/A 10187 10202 GAGCCAGAGCAGTAAA 102 2870 970157 N/A N/A 10275 10290 TACCGATGCTTCAAGA 103 2871 970167 N/A N/A 10499 10514 CACAAAGCGGGTTCAC 112 2872 970177 N/A N/A 10777 10792 AGAGAGTCCGACGCAC 118 2873 970187 N/A N/A 11006 11021 TCCCATCGAATCTGCC 105 2874 970196 N/A N/A 11136 11151 CGGTCAGCAGAACCAC 108 2875 970206 N/A N/A 11377 11392 ATGAGGGACCAGAACT 94 2876 970216 N/A N/A 11470 11485 ATCAGATTGAATCCAT 77 2877 970226 N/A N/A 11555 11570 AAGCCTAGAGGTGCCT 71 2878 970236 N/A N/A 11813 11828 CCTCATTCACAACTAG 95 2879 970246 N/A N/A 11963 11978 CTACTCTTCAGGCACC 82 2880 970256 N/A N/A 12055 12070 GGCCATTCGAGGTGAT 99 2881 970266 N/A N/A 12229 12244 GCTCATGGCTGATAAT 117 2882 970276 N/A N/A 12385 12400 TCTTTAAAATATCGCC 145 2883 970286 N/A N/A 12697 12712 ACATTAAAGGGTAAGC 123 2884 970296 N/A N/A 13042 13057 AGAACTTGCTAAGTCT 134 2885 970306 N/A N/A 13207 13222 GGTCAACACTTGGAGA 123 2886 970316 N/A N/A 13771 13786 TGCCTTACCTGACAAG 96 2887 970326 N/A N/A 14023 14038 TTTAAGAGGCGACTGC 200 2888 970336 N/A N/A 14211 14226 AAACCAGTACAGTCCA 123 2889 970346 N/A N/A 14431 14446 CCCACGCGGGAGGCTC 123 2890 970356 N/A N/A 14706 14721 CTTTGGGCACCAAAAG 133 2891 970366 N/A N/A 15411 15426 TTCATTAGAAAGCCCT 122 2892 970376 N/A N/A 15606 15621 TAGTAAGTTCTAGTCG 113 2893 970386 N/A N/A 15878 15893 CTGAGACTACACCTAC 140 2894 970396 N/A N/A 15991 16006 TTCTAGCTAACTTACA 118 2895 970406 N/A N/A 16271 16286 GACTGGAACATTGTTG 141 2896 970416 N/A N/A 16639 16654 TGCCATGGACAAGTTT 118 2897 970426 N/A N/A 16717 16732 CAAAAGGTAGTAGACC 89 2898 970436 N/A N/A 16891 16906 GCACTCTTACTCCGCT 98 2899 970446 N/A N/A 16970 16985 GAAACCCTCATCACCG 138 2900

TABLE 47 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT 23 195 969848 N/A N/A 4047 4062 CGTCGCAGGAGCGCGG 99 2901 969858 N/A N/A 4313 4328 GCACGCAAGGCTCGGA 93 2902 969868 N/A N/A 4446 4461 GGATTCCGCGCGCAGA 79 2903 969878 N/A N/A 4677 4692 CGCGACTCTGTCAGTT 92 2904 969888 N/A N/A 4981 4996 CTGCGAAGCGCGCGCG 106 2905 969898 N/A N/A 5600 5615 GCCTTCAGCGGTTTCC 96 2906 969908 N/A N/A 5719 5734 ACGGAGGCGGCAGACG 109 2907 969918 N/A N/A 5785 5800 GGTGAGAGACGAAAAC 116 2908 969928 N/A N/A 6115 6130 CCGGAGGAAGTCCCCT 100 2909 969938 N/A N/A 6277 6292 GTAGAGTGAGATGCGG 49 2910 969948 N/A N/A 6397 6412 TTTACACCGTTGCTCA 97 2911 969958 N/A N/A 6418 6433 ATGGCGCATAGGTCCT 105 2912 969968 450 465 6929 6944 CCGCTCAACCAGTTCC 112 2913 969978 N/A N/A 7011 7026 ATTCAGGAGAGCCCCT 115 2914 969988 N/A N/A 7193 7208 CTCCTATGCTGAACCC 91 2915 969998 N/A N/A 7227 7242 CCCATATACATTGAGG 84 2916 970008 N/A N/A 7603 7618 ACAGATGGCATGCGAA 81 2917 970018 N/A N/A 7666 7681 TGCTATTAAACTGATT 103 2918 970028 N/A N/A 8138 8153 CGGACACGCCCGACCC 93 2919 970038 N/A N/A 8331 8346 ACCAAAAGTACCACAG 108 2920 970048 N/A N/A 8445 8460 GACTGGAGTGAACCCT 62 2921 970058 N/A N/A 8522 8537 GGGTGCTCTTACAGCC 102 2922 970068 677 692 9167 9182 TCCGGGTGTGGCTGAT 74 2923 970078 N/A N/A 9310 9325 CCAGGATTCGCCATGG 87 2924 970088 N/A N/A 9445 9460 GCCTAGCATGGATCCT 104 2925 970098 N/A N/A 9605 9620 CACTATTCGAGAGAAA 70 2926 970108 N/A N/A 9754 9769 GGACCACATCTAATCC 112 2927 970118 N/A N/A 9852 9867 CCTGAGTTACCGCATT 76 2928 970128 N/A N/A 10011 10026 CACCTCTTCACTACCT 100 2929 970138 N/A N/A 10138 10153 TAGCCAATTCCTCCAC 99 2930 970148 N/A N/A 10193 10208 TCCATAGAGCCAGAGC 80 2931 970158 N/A N/A 10277 10292 GTTACCGATGCTTCAA 54 2932 970168 N/A N/A 10550 10565 ACTAACAGGGAGACTG 119 2933 970178 N/A N/A 10779 10794 ACAGAGAGTCCGACGC 108 2934 970188 N/A N/A 11012 11027 CTAAAGTCCCATCGAA 102 2935 970197 N/A N/A 11142 11157 CTGAAACGGTCAGCAG 97 2936 970207 N/A N/A 11399 11414 TAGGCACATCAATGTT 88 2937 970217 N/A N/A 11525 11540 AAGATCTCCATGGTGC 61 2938 970227 N/A N/A 11557 11572 CCAAGCCTAGAGGTGC 79 2939 970237 N/A N/A 11820 11835 GTGCAAGCCTCATTCA 81 2940 970247 N/A N/A 11993 12008 GTTGCCGAGATATAAA 87 2941 970257 N/A N/A 12085 12100 AGACAGTGCGCCCCAA 117 2942 970267 N/A N/A 12231 12246 GTGCTCATGGCTGATA 76 2943 970277 N/A N/A 12455 12470 TAAAACTGCGCTCTCT 114 2944 970287 N/A N/A 12860 12875 CCACATACCTGAAACG 109 2945 970297 N/A N/A 13056 13071 GAGCATCACTATTAAG 86 2946 970307 N/A N/A 13216 13231 CATGAGCTCGGTCAAC 101 2947 970317 N/A N/A 13779 13794 TAACGAGGTGCCTTAC 87 2948 970327 N/A N/A 14026 14041 TGTTTTAAGAGGCGAC 97 2949 970337 N/A N/A 14217 14232 TGTGCAAAACCAGTAC 120 2950 970347 N/A N/A 14448 14463 GGGTAGAGCAGCTCCC 116 2951 970357 N/A N/A 14710 14725 TTTGCTTTGGGCACCA 76 2952 970367 N/A N/A 15431 15446 TGCCAGTTCCTTGTGA 102 2953 970377 N/A N/A 15607 15622 ATAGTAAGTTCTAGTC 89 2954 970387 N/A N/A 15880 15895 ATCTGAGACTACACCT 108 2955 970397 N/A N/A 16095 16110 ATACACACCCTCGGGC 110 2956 970407 N/A N/A 16274 16289 ACGGACTGGAACATTG 62 2957 970417 N/A N/A 16640 16655 ATGCCATGGACAAGTT 82 2958 970427 N/A N/A 16728 16743 CCACGAAGATTCAAAA 107 2959 970437 N/A N/A 16894 16909 TGAGCACTCTTACTCC 106 2960 970447 N/A N/A 16976 16991 TGTTCAGAAACCCTCA 115 2961

TABLE 48 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  27  195 969849 N/A N/A  4081  4096 CGGTTAGCTCATCCCG  95 2962 969859 N/A N/A  4320  4335 GGCCACCGCACGCAAG 124 2963 969869 N/A N/A  4488  4503 CGACAAGTGGCGCAGA  98 2964 969879 N/A N/A  4805  4820 CGACACGCGCCGCTCG  62 2965 969889 N/A N/A  4989  5004 CTTTGAGGCTGCGAAG  98 2966 969899 N/A N/A  5610  5625 GCCCGGCCGGGCCTTC 104 2967 969909 N/A N/A  5725  5740 CCACGGACGGAGGCGG 106 2968 969919 N/A N/A  5793  5808 AGAGACGCGGTGAGAG 120 2969 969929 N/A N/A  6123  6138 CCAAATTCCCGGAGGA 125 2970 969939 N/A N/A  6280  6295 CCGGTAGAGTGAGATG 103 2971 969949 N/A N/A  6398  6413 ATTTACACCGTTGCTC  87 2972 969959 N/A N/A  6420  6435 GAATGGCGCATAGGTC  74 2973 969969  454  469  6933  6948 GGCTCCGCTCAACCAG  91 2974 969979 N/A N/A  7026  7041 TGTTAGGTGACCCAAA  98 2975 969989 N/A N/A  7198  7213 TAATTCTCCTATGCTG  97 2976 969999 N/A N/A  7243  7258 ATTCAATGCACCCCCC  98 2977 970009 N/A N/A  7604  7619 GACAGATGGCATGCGA  84 2978 970019 N/A N/A  7727  7742 ATAACACGGTGTTGAC  79 2979 970029 N/A N/A  8140  8155 GGCGGACACGCCCGAC  93 2980 970039 N/A N/A  8343  8358 GCTAAACCTGGCACCA  93 2981 970049 N/A N/A  8451  8466 CGAAAAGACTGGAGTG  74 2982 970059 N/A N/A  8784  8799 GGGTTAAAGGAGTGCA 100 2983 970069  681  696  9171  9186 GATTTCCGGGTGTGGC  25 2984 970079 N/A N/A  9371  9386 CCCCGAGCTGCACACG 105 2985 970089 N/A N/A  9457  9472 ACGAAGGGCAGTGCCT 105 2986 970099 N/A N/A  9613  9628 GAACACACCACTATTC 120 2987 970109 N/A N/A  9772  9787 GGAACTCCCGAGGGCA  87 2988 970119 N/A N/A  9854  9869 GGCCTGAGTTACCGCA  92 2989 970129 N/A N/A 10013 10028 TACACCTCTTCACTAC 118 2990 970139 N/A N/A 10143 10158 TATGTTAGCCAATTCC  59 2991 970149 N/A N/A 10196 10211 AATTCCATAGAGCCAG  82 2992 970159 N/A N/A 10279 10294 CTGTTACCGATGCTTC  44 2993 970169 N/A N/A 10558 10573 TGTAACTGACTAACAG 131 2994 970179 N/A N/A 10783 10798 CTAGACAGAGAGTCCG  95 2995 970189 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA  58 2996 970198 N/A N/A 11143 11158 GCTGAAACGGTCAGCA  92 2997 970208 N/A N/A 11401 11416 CTTAGGCACATCAATG  92 2998 970218 N/A N/A 11527 11542 GTAAGATCTCCATGGT  83 2999 970228 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC  34 3000 970238 N/A N/A 11826 11841 AAAAAGGTGCAAGCCT  94 3001 970248 N/A N/A 11996 12011 GAGGTTGCCGAGATAT  71 3002 970258 N/A N/A 12094 12109 AAGCGAGTCAGACAGT 122 3003 970268 N/A N/A 12278 12293 CTGTATGGAACCCCAA  97 3004 970278 N/A N/A 12511 12526 ATGTAAAGTCTGCTGA  80 3005 970288 N/A N/A 12862 12877 GTCCACATACCTGAAA  92 3006 970298 N/A N/A 13059 13074 ATAGAGCATCACTATT 100 3007 970308 N/A N/A 13219 13234 CAGCATGAGCTCGGTC  78 3008 970318 N/A N/A 13786 13801 TAACAGATAACGAGGT  88 3009 970328 N/A N/A 14054 14069 TCGAGATCATAGTGCA  71 3010 970338 N/A N/A 14219 14234 CCTGTGCAAAACCAGT 110 3011 970348 N/A N/A 14459 14474 TGGCGAGTGGCGGGTA 118 3012 970358 N/A N/A 14733 14748 AAGCTTAGTTATCTGG  65 3013 970368 N/A N/A 15573 15588 AGGACACTCACAGGCG  92 3014 970378 N/A N/A 15614 15629 CAGATTAATAGTAAGT 112 3015 970388 N/A N/A 15885 15900 CCGTGATCTGAGACTA  55 3016 970398 N/A N/A 16144 16159 TGACACAGGAGCCGCT  85 3017 970408 N/A N/A 16283 16298 AGATACAAAACGGACT 115 3018 970418 N/A N/A 16642 16657 TTATGCCATGGACAAG  81 3019 970428 N/A N/A 16730 16745 ACCCACGAAGATTCAA 117 3020 970438 N/A N/A 16899 16914 AGGAATGAGCACTCTT  93 3021 970448 N/A N/A 16984 16999 AGAGACCATGTTCAGA  88 3022 970608 1440 1455 19557 19572 AGATCTGTGGTAATCT 103 3023

TABLE 49 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  19  195 969850 N/A N/A  4151  4166 GGCAGTTGTGCCGTCT 117 3024 969860 N/A N/A  4342  4357 AGGGACCGCGCCAGGC 100 3025 969870 N/A N/A  4490  4505 AACGACAAGTGGCGCA  88 3026 969880 N/A N/A  4808  4823 TCCCGACACGCGCCGC 112 3027 969890 N/A N/A  5006  5021 CCACGAGGCCCCGGAG  96 3028 969900 N/A N/A  5655  5670 GACGAACGCGCAAAAC  85 3029 969910 N/A N/A  5747  5762 GCACGGAGAGGGCGAG  96 3030 969920 N/A N/A  5795  5810 ACAGAGACGCGGTGAG  86 3031 969930 N/A N/A  6211  6226 GGACTAAGGACAGCTG  87 3032 969940 N/A N/A  6283  6298 TAACCGGTAGAGTGAG  73 3033 969950 N/A N/A  6400  6415 AGATTTACACCGTTGC  70 3034 969960 N/A N/A  6423  6438 AAAGAATGGCGCATAG  90 3035 969970  471  486  6950  6965 GTCTGAGATGTCCAGC  93 3036 969980 N/A N/A  7156  7171 CCCACATAACTCAGGC 116 3037 969990 N/A N/A  7201  7216 GATTAATTCTCCTATG 119 3038 970000 N/A N/A  7375  7390 TGAGATATTCCTCTCA  82 3039 970010 N/A N/A  7628  7643 TACAGGACAGGTAAAG 129 3040 970020 N/A N/A  7735  7750 TAGAATGCATAACACG  92 3041 970030 N/A N/A  8142  8157 CAGGCGGACACGCCCG 105 3042 970040 N/A N/A  8346  8361 ATGGCTAAACCTGGCA  78 3043 970050 N/A N/A  8453  8468 TCCGAAAAGACTGGAG 107 3044 970060 N/A N/A  9095  9110 TGCTAAAGGAGTGCAG 117 3045 970070  688  703  9178  9193 GGTACGGGATTTCCGG  88 3046 970080 N/A N/A  9390  9405 AGTGAGAAAACCCCCC  92 3047 970090 N/A N/A  9459  9474 CCACGAAGGGCAGTGC 100 3048 970100 N/A N/A  9649  9664 TTTAGTATCACCTCTA  70 3049 970110 N/A N/A  9795  9810 AGGGAGCTCATTTTGA 108 3050 970120 N/A N/A  9857  9872 GAAGGCCTGAGTTACC  71 3051 970130 N/A N/A 10022 10037 AAAGAGTGGTACACCT  90 3052 970140 N/A N/A 10154 10169 GGAGCTAGTTTTATGT 128 3053 970150 N/A N/A 10201 10216 TTACTAATTCCATAGA  97 3054 970160 N/A N/A 10280 10295 ACTGTTACCGATGCTT  57 3055 970170 N/A N/A 10561 10576 CTCTGTAACTGACTAA  84 3056 970180 N/A N/A 10827 10842 TGTCACCTGGCAACCT 104 3057 970190 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC  97 3058 970199 N/A N/A 11144 11159 GGCTGAAACGGTCAGC 127 3059 970209 N/A N/A 11404 11419 ACTCTTAGGCACATCA  73 3060 970219 N/A N/A 11529 11544 AGGTAAGATCTCCATG  68 3061 970229 N/A N/A 11654 11669 CCTTAAGCTATTTGGT 886 3062 970239 N/A N/A 11848 11863 GTTACTCACGAGCACC 117 3063 970249 N/A N/A 11998 12013 ACGAGGTTGCCGAGAT  38 3064 970259 N/A N/A 12098 12113 CTGAAAGCGAGTCAGA  94 3065 970269 N/A N/A 12279 12294 ACTGTATGGAACCCCA  70 3066 970279 N/A N/A 12513 12528 TCATGTAAAGTCTGCT  79 3067 970289 N/A N/A 12876 12891 AAAGAAGCAAGTCGGT 102 3068 970299 N/A N/A 13062 13077 ATTATAGAGCATCACT 106 3069 970309 N/A N/A 13308 13323 CCACATGTCCCGTGGG 128 3070 970319 N/A N/A 13788 13803 TCTAACAGATAACGAG 103 3071 970329 N/A N/A 14089 14104 CCTGAAAAGAGCCGCC 103 3072 970339 N/A N/A 14227 14242 TAACTTCTCCTGTGCA  85 3073 970349 N/A N/A 14644 14659 GTACGAGCACATGTCA 100 3074 970359 N/A N/A 14741 14756 CTGGCCTGAAGCTTAG  75 3075 970369 N/A N/A 15587 15602 AAGTCTACAGCCCCAG 105 3076 970379 N/A N/A 15668 15683 GCACAGCCCTTGGTTA  97 3077 970389 N/A N/A 15890 15905 CACTGCCGTGATCTGA  91 3078 970399 N/A N/A 16153 16168 GAAACGAGTTGACACA 100 3079 970409 N/A N/A 16333 16348 CGCTTGTGGATATACA 100 3080 970419 N/A N/A 16650 16665 AGACCAATTTATGCCA  93 3081 970429 N/A N/A 16739 16754 CAGCATTGGACCCACG  96 3082 970439 N/A N/A 16901 16916 AGAGGAATGAGCACTC  95 3083 970449 N/A N/A 17002 17017 TGTAGAAGCCCACAAG 107 3084 970609 1443 1458 19560 19575 GATAGATCTGTGGTAA  78 3085

TABLE 50 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  21  195 969851 N/A N/A  4214  4229 GGAACGAAGAGCAGGG  88 3086 969861 N/A N/A  4344  4359 GCAGGGACCGCGCCAG  90 3087 969871 N/A N/A  4493  4508 GCAAACGACAAGTGGC 115 3088 969881 N/A N/A  4823  4838 CCGCAGCCCAAAGGCT  96 3089 969891 N/A N/A  5046  5061 CGCACTCCGGGCACCC  89 3090 969901 N/A N/A  5658  5673 ACGGACGAACGCGCAA 107 3091 969911 N/A N/A  5751  5766 GGACGCACGGAGAGGG  85 3092 969921 N/A N/A  5799  5814 AGAAACAGAGACGCGG  89 3093 969931 N/A N/A  6213  6228 TAGGACTAAGGACAGC 105 3094 969941 N/A N/A  6288  6303 ATTTATAACCGGTAGA  72 3095 969951 N/A N/A  6401  6416 CAGATTTACACCGTTG  85 3096 969961 N/A N/A  6540  6555 GCATAGGCATCCTTCC  92 3097 969971  474  489  6953  6968 CGGGTCTGAGATGTCC 102 3098 969981 N/A N/A  7160  7175 GTGACCCACATAACTC  93 3099 969991 N/A N/A  7205  7220 GTGTGATTAATTCTCC  36 3100 970001 N/A N/A  7377  7392 GATGAGATATTCCTCT 124 3101 970011 N/A N/A  7630  7645 TGTACAGGACAGGTAA  92 3102 970021 N/A N/A  7783  7798 ACCTAACTAAATGTCA 124 3103 970031 N/A N/A  8149  8164 ATTCCAACAGGCGGAC  98 3104 970041 N/A N/A  8348  8363 ATATGGCTAAACCTGG  82 3105 970051 N/A N/A  8454  8469 ATCCGAAAAGACTGGA 120 3106 970061 N/A N/A  9103  9118 TGTGAACCTGCTAAAG 110 3107 970071  690  705  9180  9195 TTGGTACGGGATTTCC  66 3108 970081 N/A N/A  9412  9427 TGGGATGCCACCATCC 102 3109 970091 N/A N/A  9466  9481 TAAGATCCCACGAAGG  91 3110 970101 N/A N/A  9659  9674 CCGTTCCTTTTTTAGT 103 3111 970111 N/A N/A  9834  9849 TAGATTCTCCCTGCAC  82 3112 970121 N/A N/A  9914  9929 GTTCAGTGTGTTGACC  70 3113 970131 N/A N/A 10100 10115 TGCTGCAAATCCCTCT  79 3114 970141 N/A N/A 10159 10174 TTTCAGGAGCTAGTTT  92 3115 970151 N/A N/A 10206 10221 CATGGTTACTAATTCC  75 3116 970161 N/A N/A 10281 10296 TACTGTTACCGATGCT  63 3117 970171 N/A N/A 10568 10583 GCGCTCTCTCTGTAAC 107 3118 970181  763  778 10828 10843 CTGTCACCTGGCAACC  89 3119 970191 N/A N/A 11085 11100 AATCACCCTGGTCACC  99 3120 970200 N/A N/A 11334 11349 ACTTATTTGTGGCTCA  64 3121 970210 N/A N/A 11407 11422 ATTACTCTTAGGCACA  70 3122 970220 N/A N/A 11531 11546 GTAGGTAAGATCTCCA  78 3123 970230 N/A N/A 11656 11671 AACCTTAAGCTATTTG  49 3124 970240 N/A N/A 11851 11866 TCAGTTACTCACGAGC 103 3125 970250 N/A N/A 12002 12017 AATCACGAGGTTGCCG 106 3126 970260 N/A N/A 12149 12164 TTTGAGACTTAACCCA  69 3127 970270 N/A N/A 12281 12296 TAACTGTATGGAACCC  72 3128 970280 N/A N/A 12590 12605 CTGGATATGTGGTGTT  70 3129 970290 N/A N/A 12892 12907 TATAACGGTGTTTCAG  64 3130 970300 N/A N/A 13066 13081 TCCTATTATAGAGCAT  97 3131 970310 N/A N/A 13352 13367 AACGACTCCACAGAGC  96 3132 970320 N/A N/A 13880 13895 GCCGAAGTCAACAGGA 117 3133 970330 N/A N/A 14115 14130 TCCAACCTTTATGATT 103 3134 970340 N/A N/A 14229 14244 TATAACTTCTCCTGTG  93 3135 970350 N/A N/A 14647 14662 GAAGTACGAGCACATG  82 3136 970360 N/A N/A 14988 15003 CAGCACCGTGGTGCGA 127 3137 970370 N/A N/A 15589 15604 GCAAGTCTACAGCCCC  51 3138 970380 N/A N/A 15801 15816 ATAACATGAGAGTGTT 117 3139 970390 N/A N/A 15892 15907 CACACTGCCGTGATCT  80 3140 970400 N/A N/A 16156 16171 AAAGAAACGAGTTGAC  86 3141 970410 N/A N/A 16379 16394 AGTAACTTGACTTGAG 103 3142 970420 N/A N/A 16658 16673 ATGGCAAAAGACCAAT 105 3143 970430 N/A N/A 16746 16761 TGCTTTGCAGCATTGG  70 3144 970440 N/A N/A 16927 16942 AGCTTACTGTGATTCT  87 3145 970450 1250 1265 17043 17058 CTTGGCAGGGAGCGGC  75 3146 970610 1445 1460 19562 19577 CGGATAGATCTGTGGT  54 3147

TABLE 51 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  28  195 969852 N/A N/A  4219  4234 CCTTAGGAACGAAGAG 106 3148 969862 N/A N/A  4346  4361 AGGCAGGGACCGCGCC 111 3149 969872 N/A N/A  4495  4510 CTGCAAACGACAAGTG 118 3150 969882 N/A N/A  4829  4844 ACGCACCCGCAGCCCA 110 3151 969892 N/A N/A  5052  5067 AGGCACCGCACTCCGG 113 3152 969902 N/A N/A  5661  5676 CGCACGGACGAACGCG 117 3153 969912 N/A N/A  5753  5768 GCGGACGCACGGAGAG 109 3154 969922 N/A N/A  5878  5893 GCTAGCAGGAGCGAGA 105 3155 969932 N/A N/A  6215  6230 CCTAGGACTAAGGACA 103 3156 969942 N/A N/A  6291  6306 GGTATTTATAACCGGT  90 3157 969952 N/A N/A  6402  6417 TCAGATTTACACCGTT  86 3158 969962 N/A N/A  6543  6558 GTTGCATAGGCATCCT  73 3159 969972  483  498  6962  6977 CACTTTGTACGGGTCT  89 3160 969982 N/A N/A  7164  7179 ACTTGTGACCCACATA 104 3161 969992 N/A N/A  7207  7222 CAGTGTGATTAATTCT  83 3162 970002 N/A N/A  7563  7578 CGCCAGCCAGATGTTT 115 3163 970012 N/A N/A  7632  7647 GTTGTACAGGACAGGT  67 3164 970022 N/A N/A  7792  7807 CAGGAACTGACCTAAC 107 3165 970032 N/A N/A  8152  8167 CATATTCCAACAGGCG 101 3166 970042 N/A N/A  8351  8366 GTCATATGGCTAAACC  77 3167 970052 N/A N/A  8478  8493 GACTGACAGCCGAAGC  73 3168 970062  626  641  9116  9131 GGCATCATGTAGTTGT 101 3169 970072  693  708  9183  9198 ACATTGGTACGGGATT  98 3170 970082 N/A N/A  9420  9435 CTTGGCTGTGGGATGC  79 3171 970092 N/A N/A  9469  9484 AAATAAGATCCCACGA 107 3172 970102 N/A N/A  9668  9683 GGTATTTTTCCGTTCC  70 3173 970112 N/A N/A  9836  9851 GCTAGATTCTCCCTGC 113 3174 970122 N/A N/A  9922  9937 TCCAAAGGGTTCAGTG  93 3175 970132 N/A N/A 10104 10119 CCCTTGCTGCAAATCC 104 3176 970142 N/A N/A 10173 10188 AACGCAAGTCTGAATT  97 3177 970152 N/A N/A 10209 10224 TCCCATGGTTACTAAT 110 3178 970162 N/A N/A 10286 10301 TCATTTACTGTTACCG  44 3179 970172  769  784 10604 10619 AACGAGCCAGTGCACA 101 3180 970182 N/A N/A 10834 10849 AGGTTCCTGTCACCTG 107 3181 970192 N/A N/A 11095 11110 GGTGCTCCTAAATCAC 101 3182 970201 N/A N/A 11335 11350 GACTTATTTGTGGCTC  80 3183 970211 N/A N/A 11410 11425 TGTATTACTCTTAGGC  35 3184 970221 N/A N/A 11538 11553 CTTTATAGTAGGTAAG 101 3185 970231 N/A N/A 11658 11673 CAAACCTTAAGCTATT  66 3186 970241 N/A N/A 11928 11943 CAAGACAAGGGTTTGA 103 3187 970251 N/A N/A 12003 12018 AAATCACGAGGTTGCC  79 3188 970261 N/A N/A 12208 12223 TGTATCATGCATACCA  97 3189 970271 N/A N/A 12285 12300 CTGGTAACTGTATGGA  83 3190 970281 N/A N/A 12591 12606 ACTGGATATGTGGTGT  90 3191 970291 N/A N/A 12894 12909 AATATAACGGTGTTTC  92 3192 970301 N/A N/A 13111 13126 GAACAAGTGTATCTTT  95 3193 970311 N/A N/A 13370 13385 GGACACCACCTCGAGG  99 3194 970321 N/A N/A 13912 13927 TCTACTGGAGTCAAGC  70 3195 970331 N/A N/A 14194 14209 TCCACCCTCCGTCTCA 109 3196 970341 N/A N/A 14231 14246 CCTATAACTTCTCCTG 107 3197 970351 N/A N/A 14650 14665 CCAGAAGTACGAGCAC 103 3198 970361 N/A N/A 14990 15005 TGCAGCACCGTGGTGC 101 3199 970371 N/A N/A 15592 15607 CGCGCAAGTCTACAGC  87 3200 970381 N/A N/A 15812 15827 TCCAACCCTAGATAAC 105 3201 970391 N/A N/A 15894 15909 TTCACACTGCCGTGAT 112 3202 970401 N/A N/A 16161 16176 CCGCAAAAGAAACGAG 107 3203 970411 N/A N/A 16386 16401 ATGGAACAGTAACTTG  97 3204 970421 N/A N/A 16666 16681 GTCACGCAATGGCAAA 102 3205 970431 N/A N/A 16770 16785 AACTAGTCGACAGCTA 107 3206 970441 N/A N/A 16933 16948 AGCAAGAGCTTACTGT 106 3207 970451 1254 1269 17047 17062 GAATCTTGGCAGGGAG 100 3208 970611 1451 1466 19568 19583 GAATGGCGGATAGATC 103 3209

TABLE 52 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ SEQ SEQ SEQ ID: 1 ID: 1 ID: 2 ID: 2 IRF4 SEQ Compound Start Stop Start Stop (% ID Number Site Site Site Site Sequence UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT  27  195 969853 N/A N/A  4221  4236 CCCCTTAGGAACGAAG 106 3210 969863 N/A N/A  4362  4377 AGCTGCGGAGCCTGGG  69 3211 969873 N/A N/A  4499  4514 GGCTCTGCAAACGACA 100 3212 969883 N/A N/A  4833  4848 TGTAACGCACCCGCAG 104 3213 969893 N/A N/A  5464  5479 GTCCGCCCCGCGCGGT  97 3214 969903 N/A N/A  5665  5680 AGAACGCACGGACGAA  99 3215 969913 N/A N/A  5756  5771 GGCGCGGACGCACGGA  90 3216 969923 N/A N/A  5996  6011 TCGCGCAAAGGGCAAG 104 3217 969933 N/A N/A  6264  6279 CGGAGGTTCCTTGAGG  58 3218 969943 N/A N/A  6293  6308 TGGGTATTTATAACCG  96 3219 969953 N/A N/A  6405  6420 CCTTCAGATTTACACC 109 3220 969963 N/A N/A  6545  6560 ATGTTGCATAGGCATC  90 3221 969973  489  504  6968  6983 CCTGTACACTTTGTAC 102 3222 969983 N/A N/A  7167  7182 TCAACTTGTGACCCAC  86 3223 969993 N/A N/A  7216  7231 TGAGGCATACAGTGTG  82 3224 970003 N/A N/A  7575  7590 CATAAAGGACCCCGCC 105 3225 970013 N/A N/A  7636  7651 CTGAGTTGTACAGGAC  45 3226 970023 N/A N/A  8100  8115 AATGGCAGCACCGTGT 100 3227 970033 N/A N/A  8216  8231 TAAGGCACTACTTCCA  90 3228 970043 N/A N/A  8382  8397 GAGATACTTGTACTGT  51 3229 970053 N/A N/A  8480  8495 TAGACTGACAGCCGAA  97 3230 970063  630  645  9120  9135 GGGTGGCATCATGTAG  73 3231 970073  699  714  9189  9204 CATGGGACATTGGTAC  82 3232 970083 N/A N/A  9429  9444 GAGAGTAAACTTGGCT  88 3233 970093 N/A N/A  9552  9567 TATTTATGAGCTTCCA  79 3234 970103 N/A N/A  9671  9686 CATGGTATTTTTCCGT  62 3235 970113 N/A N/A  9838  9853 TTGCTAGATTCTCCCT  84 3236 970123 N/A N/A  9927  9942 AAACATCCAAAGGGTT 101 3237 970133 N/A N/A 10106 10121 AGCCCTTGCTGCAAAT 111 3238 970143 N/A N/A 10176 10191 GTAAACGCAAGTCTGA  86 3239 970153 N/A N/A 10212 10227 CTTTCCCATGGTTACT  92 3240 970163 N/A N/A 10306 10321 CACCTGATCTTGCTGC  88 3241 970173 N/A N/A 10613 10628 TGATGGAGAAACGAGC 102 3242 970183  772  787 10837 10852 AAAAGGTTCCTGTCAC 105 3243 970193 N/A N/A 11100 11115 CTAATGGTGCTCCTAA  96 3244 970202 N/A N/A 11338 11353 GAGGACTTATTTGTGG  78 3245 970212 N/A N/A 11412 11427 TGTGTATTACTCTTAG  25 3246 970222 N/A N/A 11539 11554 TCTTTATAGTAGGTAA  86 3247 970232 N/A N/A 11662 11677 ATTCCAAACCTTAAGC 101 3248 970242 N/A N/A 11931 11946 GTTCAAGACAAGGGTT  85 3249 970252 N/A N/A 12006 12021 CAGAAATCACGAGGTT  75 3250 970262 N/A N/A 12211 12226 ATCTGTATCATGCATA  76 3251 970272 N/A N/A 12291 12306 CTGCCACTGGTAACTG  79 3252 970282 N/A N/A 12657 12672 GGGTGGTAGAATGTGA  67 3253 970292 N/A N/A 12926 12941 ATGAACCCTAAGTTTA 104 3254 970302 N/A N/A 13114 13129 AAGGAACAAGTGTATC  87 3255 970312 N/A N/A 13395 13410 CTCCGGAGTCAGTGCT  99 3256 970322 N/A N/A 13961 13976 GACCATCTGATCCGGA  98 3257 970332 N/A N/A 14203 14218 ACAGTCCACTCCACCC  86 3258 970342 N/A N/A 14245 14260 ACTTATAGCACTCTCC  99 3259 970352 N/A N/A 14669 14684 AAACTTGGGTCACTTA  95 3260 970362 N/A N/A 15148 15163 CTTAGAATGAGAGGTG  99 3261 970372 N/A N/A 15595 15610 AGTCGCGCAAGTCTAC 100 3262 970382 N/A N/A 15846 15861 GGCGAGTTGGCACAAT  51 3263 970392 N/A N/A 15896 15911 CATTCACACTGCCGTG  94 3264 970402 N/A N/A 16163 16178 GCCCGCAAAAGAAACG  98 3265 970412 N/A N/A 16416 16431 TCTGAGTAGACTTCTT  86 3266 970422 N/A N/A 16670 16685 CGTAGTCACGCAATGG  85 3267 970432 N/A N/A 16772 16787 GGAACTAGTCGACAGC  82 3268 970442 N/A N/A 16958 16973 ACCGAACACACCAGGT 114 3269 970452 1277 1292 17070 17085 TCTCCAAAGCATAGAG 108 3270 970612 1465 1480 19582 19597 ATTCTTGAATAGAGGA  84 3271

Example 10: Effect of Mixed MOE and cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured MM.1R cells at a density of 5,000 cells per well were transfected by free uptake with 1,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the tables below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 53 through 58 are cEt and/or MOE containing gapmers. The modified oligonucleotides have a central gap segment comprising 2′-deoxynucleosides which is flanked by wing segments on the 5′ direction and the 3′ direction. At least one nucleoside in the 5′ wing segment and/or one nucleoside in the 3′ wing segment has a MOE and/or cEt sugar modification. The “Motif” column describes the sugar modifications of each oligonucleotide. “k” indicates a cEt sugar modification; “d” indicates deoxyribose; and “e” indicates a MOE modification. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Tables 53 through 58 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 53 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID IRF4 Compound 1 Start 1 Stop 2 Start 2: Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk  13  195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk  27 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk   4 2044 1013023 N/A N/A 10142 10157 ATGTTAGCCAATTCCT k-d10-kekek 100 3272 1013024 N/A N/A 10143 10158 TATGTTAGCCAATTCC k-d10-kekek  99 2991 1013025 N/A N/A 10144 10159 TTATGTTAGCCAATTC k-d10-kekek  98 3273 1013026 N/A N/A 10145 10160 TTTATGTTAGCCAATT k-d10-kekek 100 3274 1013032 N/A N/A 10278 10293 TGTTACCGATGCTTCA k-d10-kekek  74 2294 1013033 N/A N/A 10279 10294 CTGTTACCGATGCTTC k-d10-kekek  64 2993 1013034 N/A N/A 10280 10295 ACTGTTACCGATGCTT k-d10-kekek  80 3055 1013035 N/A N/A 10281 10296 TACTGTTACCGATGCT k-d10-kekek  90 3117 1013046 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT k-d10-kekek  66 3275 1013047 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA k-d10-kekek 101 2996 1013048 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC k-d10-kekek 101 3276 1013049 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC k-d10-kekek  93 3058 1013053 N/A N/A 11119 11134 AGCAGTGATGTCAGGT k-d10-kekek  23 2718 1013054 N/A N/A 11120 11135 AAGCAGTGATGTCAGG k-d10-kekek  72 3277 1013055 N/A N/A 11121 11136 CAAGCAGTGATGTCAG k-d10-kekek  65 3278 1013060 N/A N/A 11395 11410 CACATCAATGTTTTAG k-d10-kekek  55 3279 1013061 N/A N/A 11396 11411 GCACATCAATGTTTTA k-d10-kekek  87 1159 1013062 N/A N/A 11397 11412 GGCACATCAATGTTTT k-d10-kekek  72 3280 1013063 N/A N/A 11398 11413 AGGCACATCAATGTTT k-d10-kekek  95 1233 1013074 N/A N/A 11411 11426 GTGTATTACTCTTAGG k-d10-kekek  16 1540 1013075 N/A N/A 11412 11427 TGTGTATTACTCTTAG k-d10-kekek  42 3246 1013076 N/A N/A 11413 11428 ATGTGTATTACTCTTA k-d10-kekek  83 3281 1013077 N/A N/A 11414 11429 AATGTGTATTACTCTT k-d10-kekek  76 3282 1013090 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT k-d10-kekek 101 3283 1013091 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC k-d10-kekek  60 3000 1013092 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC k-d10-kekek  70 3284 1013093 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG k-d10-kekek  53 3285 1013125 N/A N/A 12514 12529 TTCATGTAAAGTCTGC k-d10-kekek  98 3286 1013126 N/A N/A 12515 12530 GTTCATGTAAAGTCTG k-d10-kekek  55  934 1013127 N/A N/A 12516 12531 AGTTCATGTAAAGTCT k-d10-kekek  75  755 1013128 N/A N/A 12517 12532 AAGTTCATGTAAAGTC k-d10-kekek  86 3287 1013462 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-kekek  72 3272 1013463 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-kekek  92 2991 1013464 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-kekek  89 3273 1013465 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-kekek  91 3274 1013471 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-kekek  72 2294 1013472 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-kekek  48 2993 1013473 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-kekek  70 3055 1013474 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-kekek  91 3117 1013485 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-kekek  87 3275 1013486 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-kekek  74 2996 1013487 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-kekek  88 3276 1013488 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-kekek  85 3058 1013492 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-kekek  34 2718 1013493 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-kekek  57 3277 1013494 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-kekek  49 3278 1013499 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-kekek  81 3279 1013500 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-kekek  82 1159 1013501 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-kekek  77 3280 1013502 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-kekek  84 1233 1013513 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-kekek  14 1540 1013514 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-kekek  13 3246 1013515 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-kekek  46 3281 1013516 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-kekek  48 3282 1013529 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-kekek  98 3283

TABLE 54 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID IRF4 Compound 1 Start 1 Stop 2 Start 2: Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 11  195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 27 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk  3 2044 1013530 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-kekek 59 3000 1013531 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-kekek 51 3284 1013532 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-kekek 51 3285 1013564 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-kekek 90 3286 1013565 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-kekek 58  934 1013566 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-kekek 58  755 1013567 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-kekek 66 3287 1013902 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d10-keke 80 3272 1013903 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d10-keke 92 2991 1013904 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d10-keke 89 3273 1013905 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d10-keke 89 3274 1013911 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d10-keke 75 2294 1013912 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d10-keke 57 2993 1013913 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d10-keke 70 3055 1013914 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d10-keke 76 3117 1013925 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d10-keke 91 3275 1013926 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d10-keke 68 2996 1013927 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d10-keke 90 3276 1013928 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d10-keke 84 3058 1013932 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d10-keke 33 2718 1013933 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d10-keke 26 3277 1013934 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d10-keke 80 3278 1013939 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d10-keke 75 3279 1013940 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d10-keke 51 1159 1013941 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d10-keke 91 3280 1013942 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d10-keke 84 1233 1013953 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d10-keke 22 1540 1013954 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d10-keke 49 3246 1013955 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d10-keke 53 3281 1013956 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d10-keke 92 3282 1013969 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d10-keke 74 3283 1013970 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d10-keke 63 3000 1013971 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d10-keke 45 3284 1013972 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d10-keke 94 3285 1014004 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d10-keke 51 3286 1014005 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d10-keke 69  934 1014006 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d10-keke 55  755 1014007 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d10-keke 72 3287

TABLE 55 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID IRF4 Compound 1 Start 1 Stop 2 Start 2: Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk  9  195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 17 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk  2 2044 1014342 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-kdkdk 70 3272 1014343 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-kdkdk 98 2991 1014344 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-kdkdk 83 3273 1014345 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-kdkdk 89 3274 1014351 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-kdkdk 65 2294 1014352 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-kdkdk 42 2993 1014353 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-kdkdk 78 3055 1014354 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-kdkdk 93 3117 1014365 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-kdkdk 80 3275 1014366 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-kdkdk 86 2996 1014367 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-kdkdk 87 3276 1014368 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-kdkdk 97 3058 1014372 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-kdkdk 45 2718 1014373 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-kdkdk 40 3277 1014374 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-kdkdk 54 3278 1014379 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-kdkdk 72 3279 1014380 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-kdkdk 89 1159 1014381 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-kdkdk 74 3280 1014382 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-kdkdk 96 1233 1014393 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-kdkdk 11 1540 1014394 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-kdkdk 19 3246 1014395 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-kdkdk 56 3281 1014396 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-kdkdk 90 3282 1014409 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-kdkdk 77 3283 1014410 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-kdkdk 41 3000 1014411 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-kdkdk 33 3284 1014412 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-kdkdk 25 3285 1014444 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-kdkdk 86 3286 1014445 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-kdkdk 66  934 1014446 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-kdkdk 68  755 1014447 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-kdkdk 68 3287 1014783 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-eeekk 83 3272 1014784 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-eeekk 92 2991 1014785 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-eeekk 86 3273 1014786 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-eeekk 97 3274 1014792 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-eeekk 69 2294 1014793 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-eeekk 36 2993 1014794 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-eeekk 73 3055 1014795 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-eeekk 71 3117 1014806 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-eeekk 77 3275 1014807 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-eeekk 74 2996 1014808 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-eeekk 79 3276 1014809 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-eeekk 78 3058 1014813 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-eeekk 31 2718 1014814 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-eeekk 38 3277 1014815 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-eeekk 85 3278 1014820 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-eeekk 88 3279 1014821 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-eeekk 80 1159 1014822 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-eeekk 78 3280 1014823 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-eeekk 64 1233 1014834 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-eeekk 12 1540

TABLE 56 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: SEQ ID: SEQ ID: SEQ ID IRF4 Compound 1 Start 1 Stop 2 Start 2: Stop (% SEQ ID Number Site Site Site Site Sequence Motif UTC) NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 14  195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 24 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk  2 2044 1014835 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-eeekk 59 3246 1014836 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-eeekk 60 3281 1014837 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-eeekk 85 3282 1014850 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-eeekk 55 3283 1014851 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-eeekk 25 3000 1014852 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-eeekk 40 3284 1014853 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-eeekk 56 3285 1014885 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-eeekk 73 3286 1014886 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-eeekk 71  934 1014887 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-eeekk 59  755 1014888 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-eeekk 79 3287 1015224 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-ekeke 73 3272 1015225 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-ekeke 98 2991 1015226 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-ekeke 91 3273 1015227 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-ekeke 88 3274 1015233 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-ekeke 65 2294 1015234 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-ekeke 50 2993 1015235 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-ekeke 77 3055 1015236 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-ekeke 78 3117 1015247 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-ekeke 68 3275 1015248 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-ekeke 82 2996 1015249 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-ekeke 87 3276 1015250 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-ekeke 77 3058 1015254 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-ekeke 28 2718 1015255 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-ekeke 35 3277 1015256 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-ekeke 74 3278 1015261 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-ekeke 72 3279 1015262 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-ekeke 90 1159 1015263 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-ekeke 78 3280 1015264 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-ekeke 58 1233 1015275 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-ekeke 11 1540 1015276 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-ekeke 46 3246 1015277 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-ekeke 40 3281 1015278 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-ekeke 76 3282 1015291 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-ekeke 79 3283 1015292 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-ekeke 83 3000 1015293 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-ekeke 53 3284 1015294 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-ekeke 41 3285 1015326 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-ekeke 68 3286 1015327 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-ekeke 64  934 1015328 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-ekeke 49  755 1015329 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-ekeke 66 3287

TABLE 57 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 18 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 28 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 4 2044 1012810 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kkk-d10-kkk 70 3272 1012811 N/A N/A 10144 10159 TTATGTTAGCCAATTC kkk-d10-kkk 91 3273 1012812 N/A N/A 10145 10160 TTTATGTTAGCCAATT kkk-d10-kkk 75 3274 1012816 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kkk-d10-kkk 82 3275 1012817 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kkk-d10-kkk 50 3276 1012819 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kkk-d10-kkk 34 3277 1012820 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kkk-d10-kkk 40 3278 1012821 N/A N/A 11395 11410 CACATCAATGTTTTAG kkk-d10-kkk 34 3279 1012822 N/A N/A 11397 11412 GGCACATCAATGTTTT kkk-d10-kkk 82 3280 1012826 N/A N/A 11413 11428 ATGTGTATTACTCTTA kkk-d10-kkk 48 3281 1012827 N/A N/A 11414 11429 AATGTGTATTACTCTT kkk-d10-kkk 47 3282 1012835 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kkk-d10-kkk 62 3283 1012836 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kkk-d10-kkk 33 3284 1012837 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kkk-d10-kkk 46 3285 1012845 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kkk-d10-kkk 43 3286 1012846 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kkk-d10-kkk 57 3287 1015665 N/A N/A 10142 10157 ATGTTAGCCAATTCCT k-d9-kekeke 100 3272 1015666 N/A N/A 10143 10158 TATGTTAGCCAATTCC k-d9-kekeke 90 2991 1015667 N/A N/A 10144 10159 TTATGTTAGCCAATTC k-d9-kekeke 91 3273 1015668 N/A N/A 10145 10160 TTTATGTTAGCCAATT k-d9-kekeke 95 3274 1015674 N/A N/A 10278 10293 TGTTACCGATGCTTCA k-d9-kekeke 90 2294 1015675 N/A N/A 10279 10294 CTGTTACCGATGCTTC k-d9-kekeke 98 2993 1015676 N/A N/A 10280 10295 ACTGTTACCGATGCTT k-d9-kekeke 102 3055 1015677 N/A N/A 10281 10296 TACTGTTACCGATGCT k-d9-kekeke 82 3117 1015688 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT k-d9-kekeke 95 3275 1015689 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA k-d9-kekeke 112 2996 1015690 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC k-d9-kekeke 96 3276 1015691 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC k-d9-kekeke 89 3058 1015695 N/A N/A 11119 11134 AGCAGTGATGTCAGGT k-d9-kekeke 74 2718 1015696 N/A N/A 11120 11135 AAGCAGTGATGTCAGG k-d9-kekeke 66 3277 1015697 N/A N/A 11121 11136 CAAGCAGTGATGTCAG k-d9-kekeke 90 3278 1015702 N/A N/A 11395 11410 CACATCAATGTTTTAG k-d9-kekeke 102 3279 1015703 N/A N/A 11396 11411 GCACATCAATGTTTTA k-d9-kekeke 83 1159 1015704 N/A N/A 11397 11412 GGCACATCAATGTTTT k-d9-kekeke 91 3280 1015705 N/A N/A 11398 11413 AGGCACATCAATGTTT k-d9-kekeke 109 1233 1015716 N/A N/A 11411 11426 GTGTATTACTCTTAGG k-d9-kekeke 16 1540 1015717 N/A N/A 11412 11427 TGTGTATTACTCTTAG k-d9-kekeke 68 3246 1015718 N/A N/A 11413 11428 ATGTGTATTACTCTTA k-d9-kekeke 64 3281 1015719 N/A N/A 11414 11429 AATGTGTATTACTCTT k-d9-kekeke 88 3282 1015732 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT k-d9-kekeke 103 3283 1015733 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC k-d9-kekeke 94 3000 1015734 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC k-d9-kekeke 76 3284 1015735 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG k-d9-kekeke 60 3285 1015767 N/A N/A 12514 12529 TTCATGTAAAGTCTGC k-d9-kekeke 91 3286 1015768 N/A N/A 12515 12530 GTTCATGTAAAGTCTG k-d9-kekeke 76 934 1015769 N/A N/A 12516 12531 AGTTCATGTAAAGTCT k-d9-kekeke 71 755 1015770 N/A N/A 12517 12532 AAGTTCATGTAAAGTC k-d9-kekeke 86 3287

TABLE 58 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 11 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 29 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 4 2044 935595 3075 3090 21192 21207 ACTAAGCTTGATAAAG kkk-d10-kkk 86 3288 935607 4195 4210 22312 22327 AGTGTTCCAGGAGATA kkk-d10-kkk 20 3289 1012769 N/A N/A 4810 4825 GCTCCCGACACGCGCC kkk-d10-kkk 97 3290 1012772 N/A N/A 6267 6282 ATGCGGAGGTTCCTTG kkk-d10-kkk 53 3291 1012774 N/A N/A 6271 6286 TGAGATGCGGAGGTTC kkk-d10-kkk 67 3292 1012775 N/A N/A 6273 6288 AGTGAGATGCGGAGGT kkk-d10-kkk 35 3293 1012776 N/A N/A 6275 6290 AGAGTGAGATGCGGAG kkk-d10-kkk 35 3294 1012778 N/A N/A 6281 6296 ACCGGTAGAGTGAGAT kkk-d10-kkk 84 3295 1012782 N/A N/A 7634 7649 GAGTTGTACAGGACAG kkk-d10-kkk 49 3296 1012785 N/A N/A 7638 7653 GTCTGAGTTGTACAGG kkk-d10-kkk 48 3297 1012786 N/A N/A 7640 7655 AGGTCTGAGTTGTACA kkk-d10-kkk 45 3298 1012788 N/A N/A 8386 8401 AATGGAGATACTTGTA kkk-d10-kkk 74 3299 1012790 N/A N/A 8389 8404 GACAATGGAGATACTT kkk-d10-kkk 67 3300 1012791 678 693 9168 9183 TTCCGGGTGTGGCTGA kkk-d10-kkk 75 3301 1012793 680 695 9170 9185 ATTTCCGGGTGTGGCT kkk-d10-kkk 93 3302 1012795 N/A N/A 9667 9682 GTATTTTTCCGTTCCT kkk-d10-kkk 16 3303 1012796 N/A N/A 9670 9685 ATGGTATTTTTCCGTT kkk-d10-kkk 68 3304 1012799 N/A N/A 9677 9692 GTTTGCCATGGTATTT kkk-d10-kkk 59 3305 1012804 N/A N/A 9840 9855 CATTGCTAGATTCTCC kkk-d10-kkk 82 3306 1012806 N/A N/A 9846 9861 TTACCGCATTGCTAGA kkk-d10-kkk 87 3307 1012808 N/A N/A 9851 9866 CTGAGTTACCGCATTG kkk-d10-kkk 51 3308 1012809 N/A N/A 10139 10154 TTAGCCAATTCCTCCA kkk-d10-kkk 64 3309 1012813 N/A N/A 10274 10289 ACCGATGCTTCAAGAC kkk-d10-kkk 69 3310 1012814 N/A N/A 10276 10291 TTACCGATGCTTCAAG kkk-d10-kkk 84 3311 1012815 N/A N/A 10282 10297 TTACTGTTACCGATGC kkk-d10-kkk 74 3312 1012818 N/A N/A 11020 11035 GCAAGTGTCTAAAGTC kkk-d10-kkk 51 3313 1012823 N/A N/A 11400 11415 TTAGGCACATCAATGT kkk-d10-kkk 88 3314 1012825 N/A N/A 11406 11421 TTACTCTTAGGCACAT kkk-d10-kkk 56 3315 1012828 N/A N/A 11523 11538 GATCTCCATGGTGCAG kkk-d10-kkk 78 3316 1012831 N/A N/A 11530 11545 TAGGTAAGATCTCCAT kkk-d10-kkk 65 3317 1012834 N/A N/A 11564 11579 GTGCTTGCCAAGCCTA kkk-d10-kkk 73 3318 1012840 N/A N/A 11659 11674 CCAAACCTTAAGCTAT kkk-d10-kkk 82 3319 1012841 N/A N/A 11663 11678 AATTCCAAACCTTAAG kkk-d10-kkk 102 3320 1012843 N/A N/A 11995 12010 AGGTTGCCGAGATATA kkk-d10-kkk 46 3321 1012844 N/A N/A 11997 12012 CGAGGTTGCCGAGATA kkk-d10-kkk 34 3322 1012848 N/A N/A 14251 14266 GAGCCAACTTATAGCA kkk-d10-kkk 89 3323 1012850 N/A N/A 14253 14268 CAGAGCCAACTTATAG kkk-d10-kkk 70 3324 1012852 N/A N/A 14734 14749 GAAGCTTAGTTATCTG kkk-d10-kkk 55 3325 1012855 N/A N/A 14739 14754 GGCCTGAAGCTTAGTT kkk-d10-kkk 105 3326 1012859 N/A N/A 15594 15609 GTCGCGCAAGTCTACA kkk-d10-kkk 96 3327 1012860 N/A N/A 15841 15856 GTTGGCACAATTCTCT kkk-d10-kkk 78 3328 1012864 N/A N/A 15845 15860 GCGAGTTGGCACAATT kkk-d10-kkk 73 3329 1012866 N/A N/A 15848 15863 ATGGCGAGTTGGCACA kkk-d10-kkk 51 3330 1012867 N/A N/A 15850 15865 GAATGGCGAGTTGGCA kkk-d10-kkk 61 3331 1012869 N/A N/A 15884 15899 CGTGATCTGAGACTAC kkk-d10-kkk 75 3332 1012872 N/A N/A 15889 15904 ACTGCCGTGATCTGAG kkk-d10-kkk 66 3333 1012920 1441 1456 19558 19573 TAGATCTGTGGTAATC kkk-d10-kkk 80 3334 1012921 1450 1465 19567 19582 AATGGCGGATAGATCT kkk-d10-kkk 90 3335

Example 11: Effect of Mixed MOE and cEt Gapmers with Phosphorothioate Internucleoside Linkages on Human IRF4 In Vitro, Single Dose

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed and tested for their effect on IRF4 mRNA in vitro.

Cultured MM.1R cells at a density of 5,000 cells per well were transfected by free uptake with 1,000 nM concentration of modified oligonucleotide or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS4522 (forward sequence CGGAAATCCCGTACCAATGT, designated herein as SEQ ID NO: 3392; reverse sequence TGGCAACCATTTTCACAAGCT designated herein as SEQ ID NO: 3393; probe sequence TTTGGACCCCGCGGCCAC, designated herein as SEQ ID: 3394) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented in the tables below as percent control of the amount of IRF4 mRNA, relative to untreated control (UTC) cells.

The modified oligonucleotides in Tables 59 through 64 are cEt and/or MOE containing gapmers. The modified oligonucleotides have a central gap segment comprising 2′-deoxynucleosides which is flanked by wing segments on the 5′ direction and the 3′ direction. At least one nucleoside in the 5′ wing segment and/or one nucleoside in the 3′ wing segment has a MOE and/or cEt sugar modification. The “Motif” column describes the sugar modifications of each oligonucleotide. “k” indicates a cEt sugar modification; “d” indicates deoxyribose; and “e” indicates a MOE modification. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Table 59 through 64 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity. As shown below, modified oligonucleotides complementary to human IRF4 reduced the amount of human IRF4 mRNA.

TABLE 59 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 24 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 29 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 14 2044 1013023 N/A N/A 10142 10157 ATGTTAGCCAATTCCT k-d10-kekek 93 3272 1013024 N/A N/A 10143 10158 TATGTTAGCCAATTCC k-d10-kekek 99 2991 1013025 N/A N/A 10144 10159 TTATGTTAGCCAATTC k-d10-kekek 94 3273 1013026 N/A N/A 10145 10160 TTTATGTTAGCCAATT k-d10-kekek 112 3274 1013032 N/A N/A 10278 10293 TGTTACCGATGCTTCA k-d10-kekek 82 2294 1013033 N/A N/A 10279 10294 CTGTTACCGATGCTTC k-d10-kekek 74 2993 1013034 N/A N/A 10280 10295 ACTGTTACCGATGCTT k-d10-kekek 80 3055 1013035 N/A N/A 10281 10296 TACTGTTACCGATGCT k-d10-kekek 98 3117 1013046 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT k-d10-kekek 77 3275 1013047 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA k-d10-kekek 97 2996 1013048 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC k-d10-kekek 101 3276 1013049 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC k-d10-kekek 88 3058 1013053 N/A N/A 11119 11134 AGCAGTGATGTCAGGT k-d10-kekek 33 2718 1013054 N/A N/A 11120 11135 AAGCAGTGATGTCAGG k-d10-kekek 61 3277 1013055 N/A N/A 11121 11136 CAAGCAGTGATGTCAG k-d10-kekek 57 3278 1013060 N/A N/A 11395 11410 CACATCAATGTTTTAG k-d10-kekek 81 3279 1013061 N/A N/A 11396 11411 GCACATCAATGTTTTA k-d10-kekek 100 1159 1013062 N/A N/A 11397 11412 GGCACATCAATGTTTT k-d10-kekek 75 3280 1013063 N/A N/A 11398 11413 AGGCACATCAATGTTT k-d10-kekek 95 1233 1013074 N/A N/A 11411 11426 GTGTATTACTCTTAGG k-d10-kekek 20 1540 1013075 N/A N/A 11412 11427 TGTGTATTACTCTTAG k-d10-kekek 44 3246 1013076 N/A N/A 11413 11428 ATGTGTATTACTCTTA k-d10-kekek 96 3281 1013077 N/A N/A 11414 11429 AATGTGTATTACTCTT k-d10-kekek 71 3282 1013090 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT k-d10-kekek 97 3283 1013091 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC k-d10-kekek 77 3000 1013092 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC k-d10-kekek 68 3284 1013093 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG k-d10-kekek 63 3285 1013125 N/A N/A 12514 12529 TTCATGTAAAGTCTGC k-d10-kekek 118 3286 1013126 N/A N/A 12515 12530 GTTCATGTAAAGTCTG k-d10-kekek 78 934 1013127 N/A N/A 12516 12531 AGTTCATGTAAAGTCT k-d10-kekek 78 755 1013128 N/A N/A 12517 12532 AAGTTCATGTAAAGTC k-d10-kekek 94 3287 1013207 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT k-d10-kekek 77 3336 1013208 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG k-d10-kekek 53 3337 1013209 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT k-d10-kekek 91 3338 1013210 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT k-d10-kekek 66 3339 1013213 N/A N/A 18087 18102 TTATATACTGGTTGGT k-d10-kekek 78 1179 1013214 N/A N/A 18088 18103 ATTATATACTGGTTGG k-d10-kekek 78 3340 1013215 N/A N/A 18089 18104 GATTATATACTGGTTG k-d10-kekek 52 1254 1013216 N/A N/A 18090 18105 GGATTATATACTGGTT k-d10-kekek 65 1330 1013217 N/A N/A 18091 18106 GGGATTATATACTGGT k-d10-kekek 66 3341 1013218 N/A N/A 18092 18107 TGGGATTATATACTGG k-d10-kekek 57 3342 1013232 N/A N/A 18571 18586 TGATAGCTGAGCTGAT k-d10-kekek 78 2546 1013233 N/A N/A 18572 18587 GTGATAGCTGAGCTGA k-d10-kekek 71 3343 1013234 N/A N/A 18573 18588 TGTGATAGCTGAGCTG k-d10-kekek 102 3344 1013235 N/A N/A 18574 18589 ATGTGATAGCTGAGCT k-d10-kekek 103 3345 1013236 N/A N/A 18575 18590 GATGTGATAGCTGAGC k-d10-kekek 50 3346 1013237 N/A N/A 18576 18591 TGATGTGATAGCTGAG k-d10-kekek 82 3347 1013238 N/A N/A 18577 18592 TTGATGTGATAGCTGA k-d10-kekek 88 3348 1013252 N/A N/A 18611 18626 AGTGACTTGCATCCAT k-d10-kekek 98 3349 1013253 N/A N/A 18612 18627 CAGTGACTTGCATCCA k-d10-kekek 71 3350 1013254 N/A N/A 18613 18628 ACAGTGACTTGCATCC k-d10-kekek 105 3351 1013255 N/A N/A 18614 18629 GACAGTGACTTGCATC k-d10-kekek 92 3352 1013462 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-kekek 56 3272 1013463 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-kekek 97 2991 1013464 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-kekek 81 3273 1013465 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-kekek 85 3274 1013471 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-kekek 79 2294 1013472 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-kekek 56 2993 1013473 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-kekek 82 3055 1013474 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-kekek 73 3117 1013485 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-kekek 61 3275 1013486 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-kekek 91 2996 1013487 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-kekek 78 3276 1013488 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-kekek 94 3058 1013492 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-kekek 33 2718 1013493 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-kekek 56 3277 1013494 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-kekek 48 3278 1013499 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-kekek 66 3279 1013500 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-kekek 85 1159 1013501 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-kekek 81 3280 1013502 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-kekek 81 1233 1013513 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-kekek 22 1540 1013514 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-kekek 21 3246 1013515 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-kekek 51 3281 1013516 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-kekek 59 3282 1013529 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-kekek 82 3283

TABLE 60 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 18 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 36 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 10 2044 1013530 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-kekek 61 3000 1013531 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-kekek 57 3284 1013532 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-kekek 56 3285 1013564 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-kekek 88 3286 1013565 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-kekek 66 934 1013566 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-kekek 69 755 1013567 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-kekek 73 3287 1013646 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kk-d9-kekek 38 3336 1013647 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG kk-d9-kekek 33 3337 1013648 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kk-d9-kekek 56 3338 1013649 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kk-d9-kekek 35 3339 1013652 N/A N/A 18087 18102 TTATATACTGGTTGGT kk-d9-kekek 50 1179 1013653 N/A N/A 18088 18103 ATTATATACTGGTTGG kk-d9-kekek 48 3340 1013654 N/A N/A 18089 18104 GATTATATACTGGTTG kk-d9-kekek 55 1254 1013655 N/A N/A 18090 18105 GGATTATATACTGGTT kk-d9-kekek 35 1330 1013656 N/A N/A 18091 18106 GGGATTATATACTGGT kk-d9-kekek 60 3341 1013657 N/A N/A 18092 18107 TGGGATTATATACTGG kk-d9-kekek 58 3342 1013672 N/A N/A 18572 18587 GTGATAGCTGAGCTGA kk-d9-kekek 61 3343 1013673 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kk-d9-kekek 83 3344 1013674 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kk-d9-kekek 76 3345 1013675 N/A N/A 18575 18590 GATGTGATAGCTGAGC kk-d9-kekek 69 3346 1013676 N/A N/A 18576 18591 TGATGTGATAGCTGAG kk-d9-kekek 102 3347 1013677 N/A N/A 18577 18592 TTGATGTGATAGCTGA kk-d9-kekek 76 3348 1013691 N/A N/A 18611 18626 AGTGACTTGCATCCAT kk-d9-kekek 73 3349 1013692 N/A N/A 18612 18627 CAGTGACTTGCATCCA kk-d9-kekek 74 3350 1013693 N/A N/A 18613 18628 ACAGTGACTTGCATCC kk-d9-kekek 68 3351 1013694 N/A N/A 18614 18629 GACAGTGACTTGCATC kk-d9-kekek 95 3352 1013902 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d10-keke 85 3272 1013903 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d10-keke 75 2991 1013904 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d10-keke 84 3273 1013905 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d10-keke 105 3274 1013911 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d10-keke 58 2294 1013912 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d10-keke 63 2993 1013913 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d10-keke 52 3055 1013914 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d10-keke 83 3117 1013925 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d10-keke 94 3275 1013926 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d10-keke 61 2996 1013927 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d10-keke 88 3276 1013928 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d10-keke 83 3058 1013932 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d10-keke 36 2718 1013933 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d10-keke 24 3277 1013934 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d10-keke 74 3278 1013939 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d10-keke 61 3279 1013940 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d10-keke 50 1159 1013941 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d10-keke 91 3280 1013942 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d10-keke 86 1233 1013953 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d10-keke 28 1540 1013954 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d10-keke 61 3246 1013955 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d10-keke 52 3281 1013956 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d10-keke 78 3282 1013969 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d10-keke 70 3283 1013970 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d10-keke 53 3000 1013971 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d10-keke 55 3284 1013972 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d10-keke 74 3285 1014004 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d10-keke 72 3286 1014005 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d10-keke 58 934 1014006 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d10-keke 51 755 1014007 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d10-keke 80 3287 1014086 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kk-d10-keke 75 3336 1014087 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG kk-d10-keke 24 3337 1014088 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kk-d10-keke 20 3338 1014089 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kk-d10-keke 73 3339 1014092 N/A N/A 18087 18102 TTATATACTGGTTGGT kk-d10-keke 53 1179 1014093 N/A N/A 18088 18103 ATTATATACTGGTTGG kk-d10-keke 68 3340 1014094 N/A N/A 18089 18104 GATTATATACTGGTTG kk-d10-keke 55 1254 1014095 N/A N/A 18090 18105 GGATTATATACTGGTT kk-d10-keke 22 1330 1014096 N/A N/A 18091 18106 GGGATTATATACTGGT kk-d10-keke 54 3341 1014097 N/A N/A 18092 18107 TGGGATTATATACTGG kk-d10-keke 32 3342 1014111 N/A N/A 18571 18586 TGATAGCTGAGCTGAT kk-d10-keke 117 2546 1014112 N/A N/A 18572 18587 GTGATAGCTGAGCTGA kk-d10-keke 65 3343 1014113 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kk-d10-keke 84 3344 1014114 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kk-d10-keke 96 3345 1014115 N/A N/A 18575 18590 GATGTGATAGCTGAGC kk-d10-keke 36 3346 1014116 N/A N/A 18576 18591 TGATGTGATAGCTGAG kk-d10-keke 46 3347 1014117 N/A N/A 18577 18592 TTGATGTGATAGCTGA kk-d10-keke 89 3348

TABLE 61 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 24 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 32 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 11 2044 1014131 N/A N/A 18611 18626 AGTGACTTGCATCCAT kk-d10-keke 50 3349 1014132 N/A N/A 18612 18627 CAGTGACTTGCATCCA kk-d10-keke 67 3350 1014133 N/A N/A 18613 18628 ACAGTGACTTGCATCC kk-d10-keke 70 3351 1014134 N/A N/A 18614 18629 GACAGTGACTTGCATC kk-d10-keke 75 3352 1014342 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-kdkdk 97 3272 1014343 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-kdkdk 66 2991 1014344 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-kdkdk 74 3273 1014345 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-kdkdk 66 3274 1014351 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-kdkdk 74 2294 1014352 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-kdkdk 48 2993 1014353 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-kdkdk 69 3055 1014354 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-kdkdk 82 3117 1014365 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-kdkdk 95 3275 1014366 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-kdkdk 105 2996 1014367 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-kdkdk 79 3276 1014368 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-kdkdk 71 3058 1014372 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-kdkdk 52 2718 1014373 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-kdkdk 39 3277 1014374 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-kdkdk 51 3278 1014379 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-kdkdk 63 3279 1014380 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-kdkdk 93 1159 1014381 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-kdkdk 84 3280 1014382 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-kdkdk 81 1233 1014393 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-kdkdk 17 1540 1014394 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-kdkdk 29 3246 1014395 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-kdkdk 68 3281 1014396 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-kdkdk 85 3282 1014409 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-kdkdk 79 3283 1014410 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-kdkdk 41 3000 1014411 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-kdkdk 47 3284 1014412 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-kdkdk 34 3285 1014444 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-kdkdk 65 3286 1014445 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-kdkdk 64 934 1014446 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-kdkdk 53 755 1014447 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-kdkdk 66 3287 1014526 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kk-d9-kdkdk 44 3336 1014527 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG kk-d9-kdkdk 36 3337 1014528 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kk-d9-kdkdk 49 3338 1014529 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kk-d9-kdkdk 45 3339 1014532 N/A N/A 18087 18102 TTATATACTGGTTGGT kk-d9-kdkdk 82 1179 1014533 N/A N/A 18088 18103 ATTATATACTGGTTGG kk-d9-kdkdk 72 3340 1014534 N/A N/A 18089 18104 GATTATATACTGGTTG kk-d9-kdkdk 65 1254 1014535 N/A N/A 18090 18105 GGATTATATACTGGTT kk-d9-kdkdk 41 1330 1014536 N/A N/A 18091 18106 GGGATTATATACTGGT kk-d9-kdkdk 59 3341 1014537 N/A N/A 18092 18107 TGGGATTATATACTGG kk-d9-kdkdk 75 3342 1014551 N/A N/A 18571 18586 TGATAGCTGAGCTGAT kk-d9-kdkdk 122 2546 1014552 N/A N/A 18572 18587 GTGATAGCTGAGCTGA kk-d9-kdkdk 66 3343 1014553 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kk-d9-kdkdk 53 3344 1014554 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kk-d9-kdkdk 73 3345 1014555 N/A N/A 18575 18590 GATGTGATAGCTGAGC kk-d9-kdkdk 66 3346 1014556 N/A N/A 18576 18591 TGATGTGATAGCTGAG kk-d9-kdkdk 86 3347 1014557 N/A N/A 18577 18592 TTGATGTGATAGCTGA kk-d9-kdkdk 64 3348 1014571 N/A N/A 18611 18626 AGTGACTTGCATCCAT kk-d9-kdkdk 78 3349 1014572 N/A N/A 18612 18627 CAGTGACTTGCATCCA kk-d9-kdkdk 64 3350 1014573 N/A N/A 18613 18628 ACAGTGACTTGCATCC kk-d9-kdkdk 82 3351 1014574 N/A N/A 18614 18629 GACAGTGACTTGCATC kk-d9-kdkdk 107 3352 1014783 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-eeekk 94 3272 1014784 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-eeekk 73 2991 1014785 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-eeekk 91 3273 1014786 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-eeekk 79 3274 1014792 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-eeekk 61 2294 1014793 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-eeekk 44 2993 1014794 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-eeekk 66 3055 1014795 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-eeekk 92 3117 1014806 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-eeekk 123 3275 1014807 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-eeekk 88 2996 1014808 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-eeekk 54 3276 1014809 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-eeekk 69 3058 1014813 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-eeekk 44 2718 1014814 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-eeekk 49 3277 1014815 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-eeekk 92 3278 1014820 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-eeekk 89 3279 1014821 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-eeekk 80 1159 1014822 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-eeekk 64 3280 1014823 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-eeekk 57 1233 1014834 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-eeekk 25 1540

TABLE 62 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 26 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 39 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 9 2044 1014835 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-eeekk 76 3246 1014836 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-eeekk 56 3281 1014837 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-eeekk 83 3282 1014850 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-eeekk 60 3283 1014851 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-eeekk 38 3000 1014852 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-eeekk 34 3284 1014853 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-eeekk 57 3285 1014885 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-eeekk 65 3286 1014886 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-eeekk 76 934 1014887 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-eeekk 70 755 1014888 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-eeekk 97 3287 1014967 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kk-d9-eeekk 76 3336 1014968 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG kk-d9-eeekk 31 3337 1014969 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kk-d9-eeekk 60 3338 1014970 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kk-d9-eeekk 50 3339 1014973 N/A N/A 18087 18102 TTATATACTGGTTGGT kk-d9-eeekk 60 1179 1014974 N/A N/A 18088 18103 ATTATATACTGGTTGG kk-d9-eeekk 63 3340 1014975 N/A N/A 18089 18104 GATTATATACTGGTTG kk-d9-eeekk 58 1254 1014976 N/A N/A 18090 18105 GGATTATATACTGGTT kk-d9-eeekk 22 1330 1014977 N/A N/A 18091 18106 GGGATTATATACTGGT kk-d9-eeekk 65 3341 1014978 N/A N/A 18092 18107 TGGGATTATATACTGG kk-d9-eeekk 46 3342 1014992 N/A N/A 18571 18586 TGATAGCTGAGCTGAT kk-d9-eeekk 102 2546 1014993 N/A N/A 18572 18587 GTGATAGCTGAGCTGA kk-d9-eeekk 87 3343 1014994 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kk-d9-eeekk 88 3344 1014995 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kk-d9-eeekk 100 3345 1014996 N/A N/A 18575 18590 GATGTGATAGCTGAGC kk-d9-eeekk 48 3346 1014997 N/A N/A 18576 18591 TGATGTGATAGCTGAG kk-d9-eeekk 53 3347 1014998 N/A N/A 18577 18592 TTGATGTGATAGCTGA kk-d9-eeekk 83 3348 1015012 N/A N/A 18611 18626 AGTGACTTGCATCCAT kk-d9-eeekk 48 3349 1015013 N/A N/A 18612 18627 CAGTGACTTGCATCCA kk-d9-eeekk 72 3350 1015014 N/A N/A 18613 18628 ACAGTGACTTGCATCC kk-d9-eeekk 73 3351 1015015 N/A N/A 18614 18629 GACAGTGACTTGCATC kk-d9-eeekk 76 3352 1015224 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kk-d9-ekeke 70 3272 1015225 N/A N/A 10143 10158 TATGTTAGCCAATTCC kk-d9-ekeke 110 2991 1015226 N/A N/A 10144 10159 TTATGTTAGCCAATTC kk-d9-ekeke 92 3273 1015227 N/A N/A 10145 10160 TTTATGTTAGCCAATT kk-d9-ekeke 113 3274 1015233 N/A N/A 10278 10293 TGTTACCGATGCTTCA kk-d9-ekeke 51 2294 1015234 N/A N/A 10279 10294 CTGTTACCGATGCTTC kk-d9-ekeke 40 2993 1015235 N/A N/A 10280 10295 ACTGTTACCGATGCTT kk-d9-ekeke 63 3055 1015236 N/A N/A 10281 10296 TACTGTTACCGATGCT kk-d9-ekeke 86 3117 1015247 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kk-d9-ekeke 77 3275 1015248 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA kk-d9-ekeke 81 2996 1015249 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kk-d9-ekeke 101 3276 1015250 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC kk-d9-ekeke 75 3058 1015254 N/A N/A 11119 11134 AGCAGTGATGTCAGGT kk-d9-ekeke 30 2718 1015255 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kk-d9-ekeke 35 3277 1015256 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kk-d9-ekeke 66 3278 1015261 N/A N/A 11395 11410 CACATCAATGTTTTAG kk-d9-ekeke 73 3279 1015262 N/A N/A 11396 11411 GCACATCAATGTTTTA kk-d9-ekeke 88 1159 1015263 N/A N/A 11397 11412 GGCACATCAATGTTTT kk-d9-ekeke 74 3280 1015264 N/A N/A 11398 11413 AGGCACATCAATGTTT kk-d9-ekeke 65 1233 1015275 N/A N/A 11411 11426 GTGTATTACTCTTAGG kk-d9-ekeke 16 1540 1015276 N/A N/A 11412 11427 TGTGTATTACTCTTAG kk-d9-ekeke 34 3246 1015277 N/A N/A 11413 11428 ATGTGTATTACTCTTA kk-d9-ekeke 39 3281 1015278 N/A N/A 11414 11429 AATGTGTATTACTCTT kk-d9-ekeke 70 3282 1015291 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kk-d9-ekeke 99 3283 1015292 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC kk-d9-ekeke 81 3000 1015293 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kk-d9-ekeke 64 3284 1015294 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kk-d9-ekeke 39 3285 1015326 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kk-d9-ekeke 61 3286 1015327 N/A N/A 12515 12530 GTTCATGTAAAGTCTG kk-d9-ekeke 68 934 1015328 N/A N/A 12516 12531 AGTTCATGTAAAGTCT kk-d9-ekeke 51 755 1015329 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kk-d9-ekeke 71 3287 1015408 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kk-d9-ekeke 77 3336 1015409 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG kk-d9-ekeke 27 3337 1015410 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kk-d9-ekeke 40 3338 1015411 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kk-d9-ekeke 66 3339 1015414 N/A N/A 18087 18102 TTATATACTGGTTGGT kk-d9-ekeke 36 1179 1015415 N/A N/A 18088 18103 ATTATATACTGGTTGG kk-d9-ekeke 57 3340 1015416 N/A N/A 18089 18104 GATTATATACTGGTTG kk-d9-ekeke 54 1254 1015417 N/A N/A 18090 18105 GGATTATATACTGGTT kk-d9-ekeke 21 1330 1015418 N/A N/A 18091 18106 GGGATTATATACTGGT kk-d9-ekeke 66 3341 1015419 N/A N/A 18092 18107 TGGGATTATATACTGG kk-d9-ekeke 37 3342 1015433 N/A N/A 18571 18586 TGATAGCTGAGCTGAT kk-d9-ekeke 85 2546 1015434 N/A N/A 18572 18587 GTGATAGCTGAGCTGA kk-d9-ekeke 76 3343 1015435 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kk-d9-ekeke 59 3344

TABLE 63 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 33 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 30 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 13 2044 1012810 N/A N/A 10142 10157 ATGTTAGCCAATTCCT kkk-d10-kkk 67 3272 1012811 N/A N/A 10144 10159 TTATGTTAGCCAATTC kkk-d10-kkk 78 3273 1012812 N/A N/A 10145 10160 TTTATGTTAGCCAATT kkk-d10-kkk 87 3274 1012816 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT kkk-d10-kkk 72 3275 1012817 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC kkk-d10-kkk 59 3276 1012819 N/A N/A 11120 11135 AAGCAGTGATGTCAGG kkk-d10-kkk 34 3277 1012820 N/A N/A 11121 11136 CAAGCAGTGATGTCAG kkk-d10-kkk 49 3278 1012821 N/A N/A 11395 11410 CACATCAATGTTTTAG kkk-d10-kkk 25 3279 1012822 N/A N/A 11397 11412 GGCACATCAATGTTTT kkk-d10-kkk 86 3280 1012826 N/A N/A 11413 11428 ATGTGTATTACTCTTA kkk-d10-kkk 51 3281 1012827 N/A N/A 11414 11429 AATGTGTATTACTCTT kkk-d10-kkk 59 3282 1012835 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT kkk-d10-kkk 51 3283 1012836 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC kkk-d10-kkk 34 3284 1012837 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG kkk-d10-kkk 45 3285 1012845 N/A N/A 12514 12529 TTCATGTAAAGTCTGC kkk-d10-kkk 45 3286 1012846 N/A N/A 12517 12532 AAGTTCATGTAAAGTC kkk-d10-kkk 59 3287 1015436 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kk-d9-ekeke 98 3345 1015437 N/A N/A 18575 18590 GATGTGATAGCTGAGC kk-d9-ekeke 47 3346 1015438 N/A N/A 18576 18591 TGATGTGATAGCTGAG kk-d9-ekeke 40 3347 1015439 N/A N/A 18577 18592 TTGATGTGATAGCTGA kk-d9-ekeke 72 3348 1015453 N/A N/A 18611 18626 AGTGACTTGCATCCAT kk-d9-ekeke 66 3349 1015454 N/A N/A 18612 18627 CAGTGACTTGCATCCA kk-d9-ekeke 66 3350 1015455 N/A N/A 18613 18628 ACAGTGACTTGCATCC kk-d9-ekeke 91 3351 1015456 N/A N/A 18614 18629 GACAGTGACTTGCATC kk-d9-ekeke 70 3352 1015665 N/A N/A 10142 10157 ATGTTAGCCAATTCCT k-d9-kekeke 95 3272 1015666 N/A N/A 10143 10158 TATGTTAGCCAATTCC k-d9-kekeke 86 2991 1015667 N/A N/A 10144 10159 TTATGTTAGCCAATTC k-d9-kekeke 98 3273 1015668 N/A N/A 10145 10160 TTTATGTTAGCCAATT k-d9-kekeke 86 3274 1015674 N/A N/A 10278 10293 TGTTACCGATGCTTCA k-d9-kekeke 87 2294 1015675 N/A N/A 10279 10294 CTGTTACCGATGCTTC k-d9-kekeke 76 2993 1015676 N/A N/A 10280 10295 ACTGTTACCGATGCTT k-d9-kekeke 83 3055 1015677 N/A N/A 10281 10296 TACTGTTACCGATGCT k-d9-kekeke 82 3117 1015688 N/A N/A 11016 11031 GTGTCTAAAGTCCCAT k-d9-kekeke 95 3275 1015689 N/A N/A 11017 11032 AGTGTCTAAAGTCCCA k-d9-kekeke 88 2996 1015690 N/A N/A 11018 11033 AAGTGTCTAAAGTCCC k-d9-kekeke 95 3276 1015691 N/A N/A 11019 11034 CAAGTGTCTAAAGTCC k-d9-kekeke 90 3058 1015695 N/A N/A 11119 11134 AGCAGTGATGTCAGGT k-d9-kekeke 64 2718 1015696 N/A N/A 11120 11135 AAGCAGTGATGTCAGG k-d9-kekeke 65 3277 1015697 N/A N/A 11121 11136 CAAGCAGTGATGTCAG k-d9-kekeke 88 3278 1015702 N/A N/A 11395 11410 CACATCAATGTTTTAG k-d9-kekeke 82 3279 1015703 N/A N/A 11396 11411 GCACATCAATGTTTTA k-d9-kekeke 85 1159 1015704 N/A N/A 11397 11412 GGCACATCAATGTTTT k-d9-kekeke 94 3280 1015705 N/A N/A 11398 11413 AGGCACATCAATGTTT k-d9-kekeke 97 1233 1015716 N/A N/A 11411 11426 GTGTATTACTCTTAGG k-d9-kekeke 23 1540 1015717 N/A N/A 11412 11427 TGTGTATTACTCTTAG k-d9-kekeke 48 3246 1015718 N/A N/A 11413 11428 ATGTGTATTACTCTTA k-d9-kekeke 51 3281 1015719 N/A N/A 11414 11429 AATGTGTATTACTCTT k-d9-kekeke 85 3282 1015732 N/A N/A 11565 11580 TGTGCTTGCCAAGCCT k-d9-kekeke 84 3283 1015733 N/A N/A 11566 11581 GTGTGCTTGCCAAGCC k-d9-kekeke 78 3000 1015734 N/A N/A 11567 11582 CGTGTGCTTGCCAAGC k-d9-kekeke 89 3284 1015735 N/A N/A 11568 11583 ACGTGTGCTTGCCAAG k-d9-kekeke 65 3285 1015767 N/A N/A 12514 12529 TTCATGTAAAGTCTGC k-d9-kekeke 97 3286 1015768 N/A N/A 12515 12530 GTTCATGTAAAGTCTG k-d9-kekeke 90 934 1015769 N/A N/A 12516 12531 AGTTCATGTAAAGTCT k-d9-kekeke 90 755 1015770 N/A N/A 12517 12532 AAGTTCATGTAAAGTC k-d9-kekeke 79 3287 1015849 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT k-d9-kekeke 82 3336 1015850 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG k-d9-kekeke 74 3337 1015851 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT k-d9-kekeke 69 3338 1015852 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT k-d9-kekeke 107 3339 1015855 N/A N/A 18087 18102 TTATATACTGGTTGGT k-d9-kekeke 80 1179 1015856 N/A N/A 18088 18103 ATTATATACTGGTTGG k-d9-kekeke 73 3340 1015857 N/A N/A 18089 18104 GATTATATACTGGTTG k-d9-kekeke 88 1254 1015858 N/A N/A 18090 18105 GGATTATATACTGGTT k-d9-kekeke 90 1330 1015859 N/A N/A 18091 18106 GGGATTATATACTGGT k-d9-kekeke 71 3341 1015860 N/A N/A 18092 18107 TGGGATTATATACTGG k-d9-kekeke 60 3342 1015874 N/A N/A 18571 18586 TGATAGCTGAGCTGAT k-d9-kekeke 83 2546 1015875 N/A N/A 18572 18587 GTGATAGCTGAGCTGA k-d9-kekeke 84 3343 1015876 N/A N/A 18573 18588 TGTGATAGCTGAGCTG k-d9-kekeke 82 3344 1015877 N/A N/A 18574 18589 ATGTGATAGCTGAGCT k-d9-kekeke 85 3345 1015878 N/A N/A 18575 18590 GATGTGATAGCTGAGC k-d9-kekeke 82 3346 1015879 N/A N/A 18576 18591 TGATGTGATAGCTGAG k-d9-kekeke 53 3347 1015880 N/A N/A 18577 18592 TTGATGTGATAGCTGA k-d9-kekeke 66 3348 1015894 N/A N/A 18611 18626 AGTGACTTGCATCCAT k-d9-kekeke 92 3349 1015895 N/A N/A 18612 18627 CAGTGACTTGCATCCA k-d9-kekeke 81 3350 1015896 N/A N/A 18613 18628 ACAGTGACTTGCATCC k-d9-kekeke 94 3351 1015897 N/A N/A 18614 18629 GACAGTGACTTGCATC k-d9-kekeke 100 3352

TABLE 64 Percent control of human IRF4 mRNA with gapmers with phosphorothioate internucleoside linkages SEQ ID: 1 SEQ ID: 1 SEQ ID: 2 SEQ ID 2: Compound Start Stop Start Stop IRF4 SEQ Number Site Site Site Site Sequence Motif (% UTC) ID NO 609408 4226 4241 22343 22358 TTGTAAATGAGTCGGT kkk-d10-kkk 23 195 881450 3252 3267 21369 21384 ACACTTTTAGAGAGGA kkk-d10-kkk 35 2111 881659 4592 4607 22709 22724 GGAAGTTTACACTGGA kkk-d10-kkk 9 2044 935595 3075 3090 21192 21207 ACTAAGCTTGATAAAG kkk-d10-kkk 95 3288 935607 4195 4210 22312 22327 AGTGTTCCAGGAGATA kkk-d10-kkk 28 3289 1012769 N/A N/A 4810 4825 GCTCCCGACACGCGCC kkk-d10-kkk 81 3290 1012772 N/A N/A 6267 6282 ATGCGGAGGTTCCTTG kkk-d10-kkk 48 3291 1012774 N/A N/A 6271 6286 TGAGATGCGGAGGTTC kkk-d10-kkk 73 3292 1012775 N/A N/A 6273 6288 AGTGAGATGCGGAGGT kkk-d10-kkk 35 3293 1012776 N/A N/A 6275 6290 AGAGTGAGATGCGGAG kkk-d10-kkk 37 3294 1012778 N/A N/A 6281 6296 ACCGGTAGAGTGAGAT kkk-d10-kkk 98 3295 1012782 N/A N/A 7634 7649 GAGTTGTACAGGACAG kkk-d10-kkk 48 3296 1012785 N/A N/A 7638 7653 GTCTGAGTTGTACAGG kkk-d10-kkk 43 3297 1012786 N/A N/A 7640 7655 AGGTCTGAGTTGTACA kkk-d10-kkk 55 3298 1012788 N/A N/A 8386 8401 AATGGAGATACTTGTA kkk-d10-kkk 74 3299 1012790 N/A N/A 8389 8404 GACAATGGAGATACTT kkk-d10-kkk 63 3300 1012795 N/A N/A 9667 9682 GTATTTTTCCGTTCCT kkk-d10-kkk 16 3303 1012796 N/A N/A 9670 9685 ATGGTATTTTTCCGTT kkk-d10-kkk 57 3304 1012799 N/A N/A 9677 9692 GTTTGCCATGGTATTT kkk-d10-kkk 50 3305 1012804 N/A N/A 9840 9855 CATTGCTAGATTCTCC kkk-d10-kkk 70 3306 1012806 N/A N/A 9846 9861 TTACCGCATTGCTAGA kkk-d10-kkk 74 3307 1012808 N/A N/A 9851 9866 CTGAGTTACCGCATTG kkk-d10-kkk 46 3308 1012809 N/A N/A 10139 10154 TTAGCCAATTCCTCCA kkk-d10-kkk 53 3309 1012813 N/A N/A 10274 10289 ACCGATGCTTCAAGAC kkk-d10-kkk 44 3310 1012814 N/A N/A 10276 10291 TTACCGATGCTTCAAG kkk-d10-kkk 81 3311 1012815 N/A N/A 10282 10297 TTACTGTTACCGATGC kkk-d10-kkk 65 3312 1012818 N/A N/A 11020 11035 GCAAGTGTCTAAAGTC kkk-d10-kkk 47 3313 1012823 N/A N/A 11400 11415 TTAGGCACATCAATGT kkk-d10-kkk 83 3314 1012825 N/A N/A 11406 11421 TTACTCTTAGGCACAT kkk-d10-kkk 55 3315 1012828 N/A N/A 11523 11538 GATCTCCATGGTGCAG kkk-d10-kkk 88 3316 1012831 N/A N/A 11530 11545 TAGGTAAGATCTCCAT kkk-d10-kkk 67 3317 1012834 N/A N/A 11564 11579 GTGCTTGCCAAGCCTA kkk-d10-kkk 64 3318 1012840 N/A N/A 11659 11674 CCAAACCTTAAGCTAT kkk-d10-kkk 66 3319 1012841 N/A N/A 11663 11678 AATTCCAAACCTTAAG kkk-d10-kkk 74 3320 1012843 N/A N/A 11995 12010 AGGTTGCCGAGATATA kkk-d10-kkk 33 3321 1012844 N/A N/A 11997 12012 CGAGGTTGCCGAGATA kkk-d10-kkk 40 3322 1012848 N/A N/A 14251 14266 GAGCCAACTTATAGCA kkk-d10-kkk 92 3323 1012850 N/A N/A 14253 14268 CAGAGCCAACTTATAG kkk-d10-kkk 57 3324 1012852 N/A N/A 14734 14749 GAAGCTTAGTTATCTG kkk-d10-kkk 61 3325 1012855 N/A N/A 14739 14754 GGCCTGAAGCTTAGTT kkk-d10-kkk 94 3326 1012859 N/A N/A 15594 15609 GTCGCGCAAGTCTACA kkk-d10-kkk 84 3327 1012860 N/A N/A 15841 15856 GTTGGCACAATTCTCT kkk-d10-kkk 84 3328 1012864 N/A N/A 15845 15860 GCGAGTTGGCACAATT kkk-d10-kkk 63 3329 1012866 N/A N/A 15848 15863 ATGGCGAGTTGGCACA kkk-d10-kkk 52 3330 1012867 N/A N/A 15850 15865 GAATGGCGAGTTGGCA kkk-d10-kkk 60 3331 1012869 N/A N/A 15884 15899 CGTGATCTGAGACTAC kkk-d10-kkk 64 3332 1012872 N/A N/A 15889 15904 ACTGCCGTGATCTGAG kkk-d10-kkk 71 3333 1012875 N/A N/A 17236 17251 TTACGCTTATTTTTCC kkk-d10-kkk 61 3353 1012879 N/A N/A 17473 17488 CAATCTTAACCTGGAG kkk-d10-kkk 60 3354 1012881 N/A N/A 18073 18088 GTTGGCTGGTCTTTGT kkk-d10-kkk 37 3336 1012882 N/A N/A 18075 18090 TGGTTGGCTGGTCTTT kkk-d10-kkk 12 3338 1012883 N/A N/A 18076 18091 TTGGTTGGCTGGTCTT kkk-d10-kkk 24 3339 1012884 N/A N/A 18083 18098 ATACTGGTTGGTTGGC kkk-d10-kkk 31 3355 1012885 N/A N/A 18250 18265 GCCGATCATCAACTTC kkk-d10-kkk 74 3356 1012888 N/A N/A 18254 18269 CCCGGCCGATCATCAA kkk-d10-kkk 77 3357 1012889 N/A N/A 18569 18584 ATAGCTGAGCTGATCA kkk-d10-kkk 70 3358 1012890 N/A N/A 18573 18588 TGTGATAGCTGAGCTG kkk-d10-kkk 53 3344 1012891 N/A N/A 18574 18589 ATGTGATAGCTGAGCT kkk-d10-kkk 74 3345 1012892 N/A N/A 18576 18591 TGATGTGATAGCTGAG kkk-d10-kkk 41 3347 1012893 N/A N/A 18577 18592 TTGATGTGATAGCTGA kkk-d10-kkk 45 3348 1012895 N/A N/A 18581 18596 TGGATTGATGTGATAG kkk-d10-kkk 52 3359 1012897 N/A N/A 18607 18622 ACTTGCATCCATGTCA kkk-d10-kkk 65 3360 1012899 N/A N/A 18610 18625 GTGACTTGCATCCATG kkk-d10-kkk 38 3361 1012900 N/A N/A 18611 18626 AGTGACTTGCATCCAT kkk-d10-kkk 33 3349 1012901 N/A N/A 18613 18628 ACAGTGACTTGCATCC kkk-d10-kkk 66 3351 1012902 N/A N/A 18614 18629 GACAGTGACTTGCATC kkk-d10-kkk 76 3352 1012903 N/A N/A 18722 18737 AAGTGGAACTCATAGG kkk-d10-kkk 75 3362 1012904 N/A N/A 19021 19036 ATCTGTATAGTTCTCA kkk-d10-kkk 45 3363 1012906 N/A N/A 19023 19038 TAATCTGTATAGTTCT kkk-d10-kkk 58 3364 1012907 1344 1359 19461 19476 TCTGGCTAGCAGAGGT kkk-d10-kkk 74 3365 1012911 1412 1427 19529 19544 ATGTGTTCTGGTAAAT kkk-d10-kkk 66 3366 1012914 1422 1437 19539 19554 TGGATTGCTGATGTGT kkk-d10-kkk 29 3367 1012915 1424 1439 19541 19556 TCTGGATTGCTGATGT kkk-d10-kkk 64 3368 1012918 1427 1442 19544 19559 TCTTCTGGATTGCTGA kkk-d10-kkk 80 3369 1012919 1429 1444 19546 19561 AATCTTCTGGATTGCT kkk-d10-kkk 86 3370 1012920 1441 1456 19558 19573 TAGATCTGTGGTAATC kkk-d10-kkk 94 3334 1012921 1450 1465 19567 19582 AATGGCGGATAGATCT kkk-d10-kkk 75 3335

Example 12: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in SK-MEL-28 cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 185 nM, 555 nM, 1,666 nM, 5,000 nM, and 15,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 65 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% LTC) Number 185 nM 555 nM 1,666 nM 5,000 nM 15,000 nM 609311 91 79 68 36 9 609312 93 91 78 42 13 609328 99 73 57 37 14 609332 99 92 62 38 15 609333 90 82 59 32 11 609334 80 71 50 30 11 609337 84 81 57 40 15 609343 87 80 56 26 11 609354 87 79 46 25 10 609357 85 80 50 29 9 609391 94 82 53 28 10 609398 81 77 39 24 8 609407 80 71 40 25 11 609408 102 73 49 19 12

TABLE 66 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 185 nM 555 nM 1,666 nM 5,000 nM 15,000 nM 609394 94 82 59 52 22 609397 86 77 57 42 24 609405 84 86 75 40 36 609408 100 92 62 33 27 609416 89 80 54 43 17 609419 84 75 66 44 19 609422 104 87 69 50 14 609530 96 99 78 71 49 609533 96 91 80 55 37 609546 96 107 84 66 25 609571 94 86 78 58 38 609591 102 95 94 50 19 609592 98 89 67 43 31 609594 86 73 68 43 26

TABLE 67 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 185 nM 555 nM 1,666 nM 5,000 nM 15,000 nM 609394 95 76 60 52 20 609397 90 74 59 45 22 609405 96 82 77 46 37 609408 99 84 62 39 27 609416 95 76 51 39 21 609419 96 73 71 41 18 609422 90 83 82 48 17 609530 104 99 81 74 45 609533 91 97 79 58 32 609546 91 88 85 63 28 609571 96 95 78 56 35 609591 110 83 76 49 21 609592 94 88 73 43 38 609594 87 84 68 45 28

TABLE 68 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 185 nM 555 nM 1,666 nM 5,000 nM 15,000 nM 609394 101 94 61 25 6 609397 99 75 46 23 13 609405 97 80 53 17 4 609408 87 88 39 15 5 609416 99 83 47 18 6 609419 88 84 54 23 9 609422 111 92 66 17 2 609530 100 92 63 37 17 609533 92 84 65 38 19 609546 83 97 72 50 11 609571 94 90 72 47 17 609591 103 88 53 24 9 609592 90 90 55 24 8 609594 82 75 52 33 28

Example 13: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in SK-MEL-28 cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 500 nM, 1,000 nM, 2,000 nM, 4,000 nM, and 8,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 69 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 500 nM 1,000 nM 2,000 nM 4,000 nM 8,000 nM 609408 70 52 46 26 29 666273 85 97 76 53 27 666333 77 78 64 42 24 666347 80 74 56 37 19 666351 79 68 59 53 24 666378 71 57 43 28 20 666392 77 64 42 37 34 666431 85 69 47 41 24 666440 69 55 38 21 27 666441 79 60 47 29 26 666442 62 50 35 21 19 666443 71 62 46 32 13 666449 53 47 31 20 19 666458 69 52 41 22 25 666471 57 50 23 23 11

TABLE 70 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 500 nM 1,000 nM 2,000 nM 4,000 nM 8,000 nM 609408 71 58 47 37 19 666475 54 42 26 22 32 666496 54 45 30 18 13 666512 76 70 49 29 36 666534 64 55 33 19 9 666575 71 53 31 24 34 666582 80 58 45 28 30 666584 88 63 58 35 21 666586 49 28 29 19 21 666587 61 44 26 14 19 666605 68 55 38 32 18 666625 71 63 45 30 17 666640 73 59 39 42 18 666645 83 64 55 37 34 666649 80 55 48 35 21

TABLE 71 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 500 nM 1,000 nM 2,000 nM 4,000 nM 8,000 nM 609408 74 57 65 47 23 666663 78 78 43 34 67 666664 77 64 43 31 62 666678 57 42 23 25 59 666681 74 59 47 28 60 666683 64 47 35 38 55 666714 81 77 54 47 75 666726 75 60 33 28 56 666727 83 65 42 31 72 666769 98 94 94 87 71 666773 83 81 71 47 36 666782 78 79 49 44 23 666787 75 66 44 32 26 666792 84 75 61 46 32 666815 81 80 47 50 32

Example 14: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in SK-MEL-28 cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 296 nM, 888 nM, 2,666 nM, and 8,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS3114 (described hereinabove in Example 1) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 72 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 96 53 25 9 1.3 881193 90 72 54 23 2.5 881218 86 70 44 38 2.9 881242 106 65 38 27 2.2 881290 77 56 44 27 1.6 881385 99 106 84 54 >8.0 881409 91 82 51 28 2.9 881434 78 59 29 14 1.2 881506 61 37 15 11 0.5 881667 80 63 41 16 1.5 881993 94 101 73 43 >8.0 882066 99 72 53 25 2.7 882325 76 55 36 31 1.5 882326 79 73 42 31 2.3 882398 83 56 32 16 1.3 882699 88 81 65 50 >8.0 882722 97 102 84 58 >8.0 882818 134 94 75 31 4.8 882898 80 66 48 26 2.1

TABLE 73 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 85 50 22 11 1.1 881411 101 70 45 22 2.3 881412 105 74 33 21 2.1 881460 71 50 26 17 0.9 881530 83 57 34 20 1.4 881577 89 71 39 20 1.9 881578 67 48 32 18 0.8 881597 83 69 49 30 2.4 881717 70 46 20 15 0.8 881718 91 62 31 20 1.6 881741 73 49 24 16 0.9 881742 64 42 22 14 0.6 881973 79 58 43 35 2.0 882352 112 59 36 16 1.9 882725 90 68 43 20 2.0 882749 94 83 49 23 2.7 882797 69 52 27 16 0.9 882819 97 82 61 28 3.5 882866 136 119 65 28 4.5

TABLE 74 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 96 45 25 15 1.3 881127 100 72 44 21 2.3 881317 74 48 33 18 1.0 881389 79 67 42 23 1.8 881413 59 43 35 20 0.6 881414 97 79 37 20 2.2 881437 91 67 36 21 1.8 881581 62 43 27 21 0.6 881671 80 66 47 41 3.1 881743 68 55 30 24 1.0 881791 81 61 34 13 1.3 882069 74 60 39 30 1.6 882070 81 60 42 33 2.0 882162 79 67 42 17 1.6 882185 73 50 24 22 1.0 882282 65 38 22 14 0.6 882305 62 43 15 19 0.5 882376 97 88 61 33 4.1 882377 82 46 22 13 1.0

TABLE 75 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 69 41 21 11 0.7 881391 100 67 36 21 2.0 881439 76 48 32 19 1.1 881511 82 73 36 26 2.0 881558 70 49 29 14 0.9 881582 61 42 21 15 0.5 881601 92 76 47 18 2.2 881648 78 68 36 21 1.6 881672 66 51 30 18 0.8 881744 95 78 43 19 2.2 881746 69 41 26 15 0.7 881975 101 91 63 51 >8.0 881999 71 53 31 26 1.1 882210 91 78 50 22 2.5 882283 79 55 29 17 1.2 882354 81 57 34 20 1.4 882379 108 86 60 25 3.4 882800 77 55 28 15 1.1 882870 69 42 23 12 0.7

TABLE 76 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 81 47 29 18 1.1 881296 84 60 57 38 3.2 881440 75 74 46 26 2.2 881441 93 68 34 26 2.0 881465 85 63 38 29 1.9 881512 82 60 38 27 1.7 881559 84 62 47 31 2.2 881747 80 57 37 26 1.5 881955 73 66 35 30 1.6 882072 82 73 51 32 2.8 882142 85 68 48 31 2.5 882214 77 46 30 13 1.0 882309 74 54 51 24 1.6 882310 81 35 31 14 0.9 882357 74 58 35 31 1.5 882495 93 76 52 33 3.2 882565 90 77 55 33 3.3 882777 96 75 35 33 2.5 882871 67 49 44 21 1.1

TABLE 77 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 83 48 25 15 1.1 881322 98 71 42 17 2.0 881371 91 70 37 22 1.9 881394 77 47 27 14 1.0 881395 77 49 26 15 1.0 881442 51 36 22 9 0.3 881514 88 75 46 21 2.2 881561 92 62 38 18 1.7 881585 76 52 34 23 1.2 881724 100 72 39 22 2.2 881748 81 61 34 20 1.5 881749 70 46 27 12 0.8 881773 88 59 33 20 1.5 882215 83 60 50 26 2.0 882311 82 63 36 21 1.6 882358 80 52 32 17 1.2 882359 91 57 34 11 1.4 882383 92 86 64 37 4.8 882429 74 52 32 11 1.0

TABLE 78 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 85 53 36 14 1.3 881133 101 88 76 63 >8.0 881204 92 79 50 26 2.7 881396 71 46 29 16 0.9 881515 90 67 44 21 2.0 881516 81 49 35 17 1.2 881588 69 52 26 18 0.9 881726 73 42 44 8 1.0 881727 66 50 35 19 0.9 881750 73 49 19 17 0.9 882077 93 78 54 37 3.7 882099 61 46 18 23 0.6 882169 101 84 51 18 2.6 882170 92 76 39 25 2.2 882313 87 60 32 18 1.5 882384 93 68 40 18 1.9 882408 84 61 41 19 1.6 882709 104 106 108 98 >8.0 882758 82 53 36 17 1.3

TABLE 79 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 83 49 25 10 1.1 881279 87 73 43 22 2.1 881399 79 49 23 24 1.1 881422 85 63 44 29 2.1 881470 109 92 54 19 3.0 881494 68 50 33 20 0.9 881495 68 46 24 17 0.8 881517 75 41 14 12 0.7 881542 76 55 31 13 1.1 881589 80 48 26 10 1.0 881607 90 64 32 19 1.6 881608 73 45 22 21 0.8 881728 58 32 17 11 0.4 881729 94 75 40 29 2.4 881752 75 47 28 14 0.9 882409 86 66 42 20 1.8 882432 101 67 34 12 1.7 882433 72 38 25 16 0.7 882806 92 35 45 34 1.7

TABLE 80 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 99 55 43 12 1.7 690521 88 58 36 21 1.6 881112 133 107 73 56 >8.0 881183 109 93 58 31 3.9 881280 94 61 40 23 1.9 881303 90 79 55 28 3.0 881304 94 86 86 37 7.9 881327 125 115 71 44 6.9 881448 82 58 36 21 1.5 881496 72 55 34 21 1.2 881543 86 67 37 24 1.8 881610 71 49 22 13 0.8 881657 83 61 46 29 2.0 881658 77 53 29 13 1.1 881753 81 70 48 20 2.0 881962 87 67 49 29 2.5 882268 110 72 50 31 3.0 882479 96 81 45 42 3.7 882761 76 67 37 25 1.6

TABLE 81 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 87 48 28 15 1.2 690522 63 32 13 11 0.4 690523 90 61 37 19 1.7 881281 95 69 49 32 2.8 881305 76 60 42 23 1.6 881425 71 53 34 21 1.1 881426 73 45 29 19 0.9 881449 65 50 28 16 0.8 881497 129 95 57 19 3.3 881498 98 71 34 16 1.9 881659 53 25 13 9 <0.3 881660 65 32 17 10 0.5 881683 74 60 37 22 1.4 881780 82 54 34 17 1.3 881963 91 66 39 33 2.3 882175 79 43 24 12 0.9 882246 69 44 29 11 0.8 882810 100 66 45 25 2.3 882833 101 86 57 23 3.1

TABLE 82 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 87 51 28 14 1.2 881258 115 91 60 31 3.9 881282 104 89 42 32 3.1 881404 89 81 43 27 2.5 881427 87 65 48 19 1.9 881450 63 42 26 12 0.6 881451 100 77 49 30 2.9 881452 77 67 49 27 2.2 881499 84 74 44 35 2.7 881661 122 92 51 23 3.2 881684 106 89 55 33 3.8 881733 82 63 32 29 1.7 881734 72 49 34 23 1.0 881941 94 86 60 41 5.2 882177 94 62 35 30 2.0 882199 90 81 38 16 2.0 882200 89 60 36 22 1.7 882247 79 60 47 33 2.2 882765 101 82 57 30 3.5

TABLE 83 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 87 49 30 11 1.2 881213 85 67 50 20 2.1 881238 83 58 33 19 1.4 881286 64 51 35 22 0.9 881381 77 55 34 26 1.3 881382 83 71 49 37 3.0 881405 86 58 42 16 1.6 881477 80 62 56 28 2.4 881478 102 69 47 30 2.7 881571 109 90 47 27 3.1 881572 87 67 39 25 1.9 881616 104 76 47 36 3.3 881663 130 108 63 32 4.6 881784 117 87 50 35 3.7 882085 82 66 46 27 2.1 882107 109 83 74 37 5.5 882178 114 69 36 16 2.1 882601 123 99 64 26 4.0 882670 91 85 66 34 4.7

TABLE 84 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 296 nM 888 nM 2,666 nM 8,000 nM (μM) 609408 101 60 32 14 1.6 881240 103 79 47 23 2.6 881359 124 89 57 35 4.1 881360 83 58 40 22 1.6 881384 85 72 42 39 2.8 881432 73 62 49 29 2.0 881575 92 67 52 40 3.4 881593 84 71 57 31 3.1 881761 94 93 72 56 >8.0 882039 95 79 68 50 >8.0 882086 96 79 50 25 2.7 882087 72 55 36 32 1.4 882204 67 77 38 30 1.9 882227 111 90 54 47 5.2 882228 96 75 52 40 3.8 882323 106 87 62 34 4.3 882532 114 85 54 33 3.7 882721 98 78 69 45 6.6 882744 84 79 50 31 2.9

Example 15: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 74 nM, 222 nM, 666 nM, and 2,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 85 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 74 nM 222 nM 666 nM 2,000 nM (μM) 609408 86 66 43 17 0.4 935580 89 62 34 10 0.4 935603 91 69 46 23 0.5 935620 84 64 37 15 0.4 935658 82 56 37 15 0.3 935898 74 47 24 7 0.2 935911 100 88 66 43 1.5 935918 90 75 45 23 0.6 935921 80 52 27 9 0.3 935925 98 67 62 36 0.9 935928 90 72 48 23 0.6 935929 96 85 60 36 1.1 935935 95 83 63 32 1.0 935939 86 74 50 41 0.9 935941 86 84 70 41 1.7 935948 86 62 45 24 0.5 935958 83 56 33 14 0.3 935961 78 51 27 8 0.3 935968 88 61 35 12 0.4

TABLE 86 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 74 nM 222 nM 666 nM 2,000 nM (μM) 609408 79 59 37 14 0.3 935581 78 61 38 15 0.4 935608 85 71 48 20 0.5 935655 93 88 70 46 >2.0 935679 92 72 47 23 0.6 935689 86 63 40 19 0.4 935696 72 45 22 9 0.2 935697 80 55 23 9 0.3 935698 83 68 39 17 0.4 935699 87 60 36 14 0.4 935700 72 59 26 7 0.3 935701 82 64 40 18 0.4 935707 84 68 49 23 0.5 935708 84 59 34 15 0.4 935721 86 70 46 24 0.5 935724 73 39 20 5 0.2 935727 85 67 49 27 0.6 935734 74 58 35 16 0.3 935741 85 68 42 17 0.4

TABLE 87 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 74 nM 222 nM 666 nM 2,000 nM (μM) 609408 77 60 41 16 0.4 935668 94 78 51 25 0.7 935671 91 73 54 27 0.7 935686 90 69 53 34 0.8 935709 72 78 57 36 0.9 935731 80 66 52 36 0.7 935762 72 56 35 19 0.3 935765 75 55 37 15 0.3 935772 81 65 51 27 0.6 935779 78 72 55 28 0.7 935782 66 55 36 15 0.2 935789 81 77 58 32 0.9 935795 95 77 60 33 0.9 935805 85 68 42 21 0.5 935850 74 48 25 12 0.2 935851 78 54 26 9 0.3 935854 79 64 35 14 0.3 935857 78 66 44 19 0.4 935878 71 42 24 10 0.2

TABLE 88 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 74 nM 222 nM 666 nM 2,000 nM (μM) 609408 79 62 39 15 0.4 881450 100 75 56 28 0.8 881659 76 45 21 5 0.2 935824 86 65 46 23 0.5 935833 81 60 44 26 0.5 935840 82 54 51 27 0.5 935853 85 64 45 21 0.5 935856 92 71 50 29 0.7 935859 90 81 63 36 1.1 935888 79 72 43 20 0.5 936006 80 64 41 21 0.4 936007 87 65 34 13 0.4 936011 94 79 62 40 1.2 936013 78 67 48 26 0.5 936016 83 62 43 21 0.4 936018 82 66 43 17 0.4 936033 80 63 47 25 0.5 936039 85 67 48 25 0.5 936046 88 66 49 30 0.6

Example 16: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 74 nM, 222 nM, 666 nM, and 2,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 89 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609311 94 106 100 87 609312 97 96 103 93 609328 87 86 88 83 609337 94 98 99 96 609354 93 88 76 50 609357 101 87 90 61 609408 83 57 35 11 609422 96 100 90 76 609530 103 94 94 87 609533 103 103 91 75 609546 100 103 107 97 609591 93 94 89 68 609592 95 94 88 76

TABLE 90 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609408 93 61 37 12 609592 97 85 88 81 609594 98 97 81 85 881127 96 98 87 66 881183 93 89 75 55 881193 25 21 17 13 881204 97 82 49 26 881955 97 73 44 16 881962 119 96 93 67 881963 104 119 104 91 881973 99 95 89 74 881999 98 91 80 46 882066 108 103 104 94 882069 109 99 91 65 882070 108 106 81 68 882072 112 104 94 72 882077 107 122 101 102 882085 96 85 71 37 882086 90 77 50 17

TABLE 91 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609408 84 68 36 12 882087 84 75 52 23 882142 95 99 82 68 882162 87 94 76 49 882169 97 79 59 41 882170 86 77 60 28 882175 98 101 72 46 882177 94 96 80 60 882178 103 105 109 95 882185 100 95 87 63 882199 98 105 98 85 882200 111 118 101 87 882204 101 96 78 61 882214 113 106 96 80 882215 112 113 110 104 882227 114 100 109 90 882228 101 92 85 63 882247 114 105 104 102 882268 108 94 89 67

TABLE 92 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609408 81 69 39 9 882283 98 94 86 54 882309 85 93 79 49 882310 91 94 85 92 882311 83 74 59 18 882313 87 73 57 25 882323 92 95 81 50 882325 99 87 62 31 882326 100 98 100 68 882352 93 91 78 62 882354 96 93 78 66 882357 103 84 78 48 882358 90 74 42 13 882359 109 112 104 91 882376 104 107 97 86 882377 98 94 89 64 882379 101 101 108 101 882383 106 112 104 104 882384 101 98 80 62

TABLE 93 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609408 74 64 38 13 882398 81 74 63 28 882408 88 77 64 37 882409 101 67 89 92 882429 98 90 90 89 882432 88 93 91 83 882479 86 91 89 56 882495 83 100 96 87 882532 89 101 126 N/A 882565 107 109 109 92 882601 103 98 85 91 882670 104 95 91 60 882721 98 97 91 73 882725 93 92 80 72 882744 100 66 73 48 882749 89 92 77 55 882758 88 79 72 56 882761 87 88 77 61 882765 89 103 94 98

TABLE 94 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 74 nM 222 nM 666 nM 2,000 nM 609408 54 34 41 N/A 881450 65 59 29 11 881659 45 19 6 2 882777 109 133 93 105 882797 113 106 93 63 882800 112 83 56 22 882806 100 115 107 91 882810 88 88 84 50

Example 17: Design of 3-10-3 cEt Gapmers with Phosphorothioate Internucleoside Linkages

Modified oligonucleotides complementary to a human IRF4 nucleic acid were designed. The modified oligonucleotides in Table 95 are 3-10-3 cEt gapmers. The gapmers are 16 nucleobases in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on both the 5′ end and on the 3′ end comprising three cEt nucleosides. The sugar motif for the gapmers is (from 5′ to 3′): kkkddddddddddkkk; wherein ‘d’ represents a 2′-deoxyribose sugar and ‘k’ represents a cEt modified sugar. Each internucleoside linkage is a phosphorothioate internucleoside linkage and each cytosine residue is a 5-methylcytosine. “Start Site” indicates the 5′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside to which the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in Table 95 below is complementary to human IRF4 nucleic acid sequences SEQ ID NO: 1 or SEQ ID NO: 2, as indicated. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular nucleic acid sequence with 100% complementarity.

TABLE 95 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides SEQ ID NO: SEQ ID NO: SEQ ID NO: SEQ ID NO: Compound 1 Start 1 Stop 2 Start 2 Stop SEQ Number Site Site Site Site Sequence ID NO 970454 N/A N/A 17700 17715 AGAACATTACGAGAGG 3371 970466 N/A N/A 18074 18089 GGTTGGCTGGTCTTTG 3337 970500 N/A N/A 18088 18103 ATTATATACTGGTTGG 3340 970524 N/A N/A 18248 18263 CGATCATCAACTTCTT 3372 970527 N/A N/A 18572 18587 GTGATAGCTGAGCTGA 3343 970539 N/A N/A 18583 18598 ACTGGATTGATGTGAT 3373 970545 N/A N/A 18718 18733 GGAACTCATAGGTGTA 3374 970546 1415 1430 19532 19547 CTGATGTGTTCTGGTA 3375 970547 N/A N/A 17235 17250 TACGCTTATTTTTCCA 3376 970548 N/A N/A 17474 17489 GCAATCTTAACCTGGA 3377 970552 N/A N/A 18580 18595 GGATTGATGTGATAGC 3378 970554 N/A N/A 19020 19035 TCTGTATAGTTCTCAA 3379 970574 1350 1365 19467 19482 TAGTTGTCTGGCTAGC 3380 970597 N/A N/A 18575 18590 GATGTGATAGCTGAGC 3346 970598 N/A N/A 18612 18627 CAGTGACTTGCATCCA 3350 970600 1346 1361 19463 19478 TGTCTGGCTAGCAGAG 3381 970602 1396 1411 19513 19528 CGTAGCCCCTCAGGAA 3382 970603 1423 1438 19540 19555 CTGGATTGCTGATGTG 3383

Example 18: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 62.5 nM, 250 nM, 1,000 nM, and 4,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 96 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 84 55 28 7 935583 99 77 32 8 969936 86 88 61 34 969937 94 100 75 33 969938 104 84 60 30 970044 115 107 79 42 970069 86 111 97 67 970104 101 87 60 30 970114 109 96 73 45 970117 109 84 48 19 970139 78 60 44 24 970158 101 102 81 46 970159 82 64 45 18 970189 85 66 42 25 970217 96 90 71 44 970228 65 77 37 13 970253 111 105 100 63 970344 83 117 124 92 970388 100 87 70 29

TABLE 97 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 92 55 28 8 969879 106 110 107 113 969933 94 71 48 36 969991 115 92 56 13 970013 96 78 48 19 970043 102 82 43 16 970103 96 64 52 34 970160 111 96 75 35 970161 106 104 82 62 970162 94 82 60 18 970211 93 79 59 32 970212 97 80 38 7 970230 110 105 84 52 970231 113 105 114 94 970249 116 93 59 22 970358 113 110 87 55 970370 107 93 77 32 970382 113 97 70 40 970610 125 103 88 50

TABLE 98 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 84 56 30 7 970454 98 84 59 35 970466 102 84 51 34 970500 97 85 61 23 970524 76 50 28 8 970527 87 62 30 11 970539 98 94 75 35 970545 89 74 47 25 970546 86 73 55 31 970547 99 71 49 35 970548 103 92 63 30 970552 93 76 41 12 970554 102 84 58 31 970574 96 84 68 32 970597 38 21 15 8 970598 60 57 47 25 970600 49 35 29 21 970602 41 39 32 19 970603 55 47 44 17

Example 19: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 62.5 nM, 250 nM, 1,000 nM, and 4,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4523 (forward sequence AAGCCTTGGCGTTCTCAGACT, designated herein as SEQ ID NO: 3386; reverse sequence TCAGCTCCTTCACGAGGATTTC, designated herein as SEQ ID NO: 3387; probe sequence CCGGCTGCACATCTGCCTGTACTACC, designated herein as SEQ ID: 3388), was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the table below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 99 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 75 51 31 10 970454 88 80 49 34 970466 85 80 65 30 970500 91 75 43 26 970524 77 50 25 7 970527 83 53 25 12 970539 91 80 61 31 970545 68 58 35 20 970546 64 59 47 30 970547 88 56 44 23 970548 97 92 62 29 970552 76 80 40 12 970554 100 74 60 29 970574 93 76 67 32 970597 93 99 78 51 970598 80 76 62 39 970600 82 87 80 63 970602 85 80 66 41 970603 83 77 64 29

Example 20: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 62.5 nM, 250 nM, 1,000 nM, and 4,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set hIRF4_LTS34726 (described hereinabove in Example 7) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 100 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 72 43 19 4 881955 89 69 40 10 881962 107 92 66 23 881963 116 116 99 88 881973 109 101 86 59 881999 117 98 72 32 882066 110 114 107 105 882069 100 95 75 43 882070 101 94 82 42 882072 95 98 83 59 882077 101 96 90 68 882085 96 71 49 13 882086 92 68 37 8 882087 98 74 45 13 882142 103 89 80 56 882162 94 80 61 27 882169 102 93 93 77 882170 99 84 71 35 882175 89 75 45 21

TABLE 101 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 73 43 19 4 882177 83 66 38 11 882178 101 100 87 59 882185 99 90 73 41 882199 108 111 96 82 882200 112 106 97 66 882204 96 82 55 22 882214 98 92 53 12 882215 118 112 117 106 882228 89 75 46 15 882246 101 98 85 58 882247 97 101 92 74 882268 101 88 78 49 882283 104 95 86 48 882309 87 76 59 31 882310 96 103 105 107 882311 99 99 119 99 882313 98 82 61 31 882323 89 89 81 68

TABLE 102 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 74 49 23 5 882325 94 95 85 45 882326 101 98 86 64 882352 97 88 75 48 882354 103 91 52 38 882357 106 91 68 36 882358 96 92 74 37 882359 98 99 94 64 882377 104 99 82 43 882384 96 82 67 39 882398 85 63 36 10 882408 91 68 41 11 882409 105 94 75 44 882429 105 99 90 74 882432 101 102 102 103 882479 98 99 94 83 882495 97 94 89 74 882532 93 92 78 41 882565 100 99 94 88

TABLE 103 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 77 50 23 6 881955 92 75 43 11 881962 96 91 64 26 881963 95 100 93 76 882725 102 102 96 78 882744 104 107 93 74 882749 102 94 79 55 882758 108 94 76 44 882761 106 99 77 47 882765 99 89 61 34 882777 105 111 108 90 882797 107 96 78 44 882800 89 68 36 9 882806 99 96 84 60 882810 94 93 65 31 882833 101 102 83 67 882870 98 88 69 34 882871 103 90 75 55 882898 101 88 72 45

Example 21: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 62.5 nM, 250 nM, 1,000 nM, and 4,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4523 (described hereinabove in Example 19) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the table below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 104 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 62.5 nM 250 nM 1,000 nM 4,000 nM 609408 84 47 22 5 881955 82 64 33 8 881962 109 79 63 25 881963 99 78 74 66 882725 85 72 76 60 882744 85 104 80 60 882749 114 95 64 50 882758 111 98 67 37 882761 114 108 75 32 882765 85 75 52 27 882777 83 95 79 63 882797 80 72 59 40 882800 53 60 28 7 882806 108 95 78 55 882810 128 119 69 34 882833 94 91 75 69 882870 114 97 71 31 882871 79 82 55 40 882898 81 73 58 34

Example 22: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in MM.1R cells. Cells were plated at a density of 5,000 cells per well and transfected by free uptake with 62.5 nM, 250 nM, 1,000 nM, and 4,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 24 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 105 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 62.5 nM 250 nM 1,000 nM 4,000 nM (μM) 609408 61 41 26 12 0.1 881450 68 59 41 26 0.4 881659 59 34 16 9 0.1 1013053 63 50 32 16 0.2 1013074 89 71 49 14 0.7 1013492 66 46 36 21 0.2 1013513 72 64 37 21 0.4 1013514 81 68 41 15 0.5 1013647 66 56 37 18 0.3 1013933 92 78 44 13 0.7 1013953 97 62 40 13 0.6 1014087 82 67 40 28 0.7 1014088 59 44 32 15 0.2 1014095 42 35 24 12 <0.1 1014097 79 57 40 17 0.4 1014393 60 49 26 9 0.2 1014394 46 38 28 9 <0.1 1014412 85 57 44 23 0.6 1014834 50 40 22 9 <0.1

TABLE 106 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) IC₅₀ Number 62.5 nM 250 nM 1,000 nM 4,000 nM (μM) 609408 63 46 28 15 0.2 881659 58 32 16 9 0.1 935607 57 41 28 16 0.1 1012795 66 43 26 7 0.2 1012819 92 75 35 25 0.7 1012821 52 38 28 11 0.1 1012836 103 69 44 19 0.8 1012882 94 60 31 11 0.5 1012883 60 42 28 11 0.1 1012884 49 39 25 14 <0.1 1012900 90 72 42 22 0.7 1012914 55 42 25 10 0.1 1014968 56 48 35 16 0.2 1014976 76 43 37 14 0.3 1015254 68 46 31 14 0.2 1015275 86 61 24 12 0.4 1015409 66 53 34 16 0.3 1015417 76 56 37 16 0.4 1015716 59 46 24 10 0.1

Example 23: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in KMS11 cells for their effects on target knockdown and on cell line proliferation.

Target Knockdown

KMS11 cells were plated at a density of 10,000 cells per well and transfected by free uptake with 8 nM, 40 nM, 200 nM, and 1,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 48 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 107 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 79 67 38 14 935762 79 41 17 10 935918 71 35 14 10 936007 81 67 25 14 970527 90 53 23 15 882085 86 56 24 11 882408 74 70 37 17

TABLE 108 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 92 69 30 13 882800 78 52 24 10 969933 96 88 72 61 1012795 70 41 18 9 1012821 84 53 39 20 1012884 85 51 26 17 1014095 82 41 19 10

TABLE 109 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 89 67 30 14 1014393 80 50 22 12 1014394 86 74 38 19 1014834 86 51 17 9 1015716 95 78 43 17 881413 92 64 35 13 881449 94 85 48 22

TABLE 110 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 98 78 33 15 935658 87 61 23 13 935696 90 48 17 11 935898 103 81 50 31 935928 92 48 17 11 935968 95 52 17 11 936006 99 81 38 19 Proliferation KMS11 cells were plated at a density of 2,000 cells per well and transfected by free uptake with 8 nM, 40 nM, 200 nM, and 1,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After seven days, CellTiterGlo-2.0 (Promega) was added and luminescence was measured on Glomax (Promega).

TABLE 111 Dose-dependent reduction of KMS11 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 93 79 22 7 935762 89 34 9 6 935918 85 28 8 7 936007 93 66 20 8 970527 92 50 11 5 882085 97 55 11 6 882408 95 68 25 10

TABLE 112 Dose-dependent reduction of KMS11 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 98 82 26 9 882800 87 48 16 9 969933 93 86 79 78 1012795 89 25 4 1 1012821 98 47 22 5 1012884 104 61 24 12 1014095 95 35 15 13

TABLE 113 Dose-dependent reduction of KMS11 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 91 67 21 7 1014393 89 45 15 9 1014394 91 70 26 11 1014834 86 43 12 6 1015716 92 78 32 12 881413 98 67 15 11 881449 97 81 39 21

TABLE 114 Dose-dependent reduction of KMS11 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 90 77 22 8 935658 91 51 11 7 935696 86 29 7 6 935898 96 82 56 38 935928 94 45 9 7 935968 94 49 10 5 936006 97 80 26 12

Example 24: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in H929 cells for their effects on target knockdown and on cell line proliferation.

Target Knockdown

H929 cells were plated at a density of 10,000 cells per well and transfected by free uptake with 8 nM, 40 nM, 200 nM, and 1,000 nM concentrations of modified oligonucleotide or 0.67 nM, 2 nM, 6.67 nM or 20 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 48 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to that of untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 115 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 88 66 39 34 935762 76 47 29 31 935918 70 43 34 24 936007 85 62 39 32 970527 83 49 41 24 882085 107 70 48 16 882408 99 88 59 42

TABLE 116 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 69 53 35 30 882800 63 62 38 28 969933 112 95 87 83 1012795 72 67 45 31 1012821 69 86 47 35 1012884 82 88 51 32 1014095 71 68 31 21

TABLE 117 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 609408 88 57 31 37 1014393 70 43 38 24 1014394 83 60 43 32 1014834 84 45 33 23 1015716 88 75 46 17 881413 86 67 46 24 881449 109 90 63 51

TABLE 118 Dose-dependent reduction of human IRF4 mRNA by modified oligonucleotides Compound IRF4 expression (% UTC) Number 0.67 nM 2 nM 6.67 nM 20 nM 609408 74 53 26 37 935658 84 75 48 32 935696 78 59 33 26 935898 94 79 55 43 935928 74 54 36 28 935968 99 73 55 40 936006 91 74 61 52 Proliferation H929 cells were plated at a density of 2,000 cells per well and transfected by free uptake with 8 nM, 40 nM, 200 nM, and 1,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After seven days, CellTiterGlo-2.0 (Promega) was added and luminescence was measured on Glomax (Promega).

TABLE 119 Dose-dependent reduction of H929 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 94 69 32 12 935762 71 27 16 3 935918 87 58 26 2 936007 89 64 26 3 970527 95 58 15 1 882085 96 65 24 1 882408 88 65 45 9

TABLE 120 Dose-dependent reduction of H929 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 73 75 21 6 882800 89 87 30 4 969933 92 90 77 77 1012795 89 86 36 4 1012821 95 86 41 13 1012884 68 62 9 2 1014095 78 63 22 5

TABLE 121 Dose-dependent reduction of H929 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 86 74 48 30 1014393 95 65 30 2 1014394 77 53 35 6 1014834 89 61 34 2 1015716 99 74 31 2 881413 85 53 33 4 881449 94 76 54 25

TABLE 122 Dose-dependent reduction of H929 proliferation by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 8 nM 40 nM 200 nM 1,000 nM 690890 92 79 52 30 935658 95 62 23 7 935696 84 60 32 3 935898 101 87 62 26 935928 75 50 25 6 935968 96 66 40 10 936006 98 73 38 9

Example 25: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in ABC-DLBCL lines U2932 and TMD8 for their effects on target knockdown and on cell line proliferation.

Target Knockdown

Cells were plated at a density of 10,000 cells per well and transfected by free uptake with 50 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. Control oligonucleotide ION 792169, a 3-10-3 cEt gapmer with the sequence CGCCGATAAGGTACAC (SEQ ID NO: 3384), was also included. After a treatment period of approximately 48 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 123 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in U2932 cells Compound IRF4 expression (% UTC) Number 50 nM 200 nM 1,000 nM 5,000 nM 792169 98 102 102 99 690890 84 64 55 38 882800 85 63 51 38 695696 71 62 54 49 695968 84 70 55 43 695918 83 64 47 33

TABLE 124 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in TMD8 cells Compound IRF4 expression (% UTC) Number 50 nM 200 nM 1,000 nM 5,000 nM 792169 115 99 99 91 690890 97 75 53 26 882800 97 64 38 19 695696 94 68 45 29 695968 95 63 46 25 695918 105 65 44 24 Proliferation Cells were plated at a density of 2,000 cells per well and transfected by free uptake with 50 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After seven days, CellTiterGlo-2.0 (Promega) was added and luminescence was measured on Glomax (Promega).

TABLE 125 Dose-dependent reduction of proliferation of U2932 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 50 nM 200 nM 1,000 nM 5,000 nM 792169 99 115 111 75 690890 111 109 63 2 882800 113 103 40 2 695696 107 111 77 17 695968 102 101 74 24 695918 89 100 70 7

TABLE 126 Dose-dependent reduction of proliferation of TMD8 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 50 nM 200 nM 1,000 nM 5,000 nM 792169 93 108 102 98 690890 110 120 118 17 882800 132 149 108 27 695696 125 143 94 5 695968 85 131 131 10 695918 139 130 122 4

Example 26: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in ALCL cell lines for their effects on target knockdown and on cell line proliferation.

Target Knockdown

Cells were plated at a density of 10,000 cells per well and transfected by free uptake with 16 nM, 80 nM, or 400 nM concentrations of modified oligonucleotide, or 40 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After a treatment period of approximately 48 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to untreated control (UTC) cells. Control oligonucleotide 549148, a 3-10-3 cEt gapmer with the sequence GGCTACTACGCCGTCA (SEQ ID NO: 3385), was also included. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 127 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in Karpas299 cells Compound IRF4 expression (% UTC) Number 16 nM 80 nM 400 nM 2,000 nM 549148 92 85 86 83 609408 85 65 39 23 609416 82 75 66 53

TABLE 128 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in SupM2 cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 109 100 118 117 609408 83 91 32 10 609416 93 85 48 21 Proliferation Cells were plated at a density of 2,000 cells per well and transfected by free uptake with 50 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After seven days, CellTiterGlo-2.0 (Promega) was added and luminescence was measured on Glomax (Promega).

TABLE 129 Dose-dependent reduction of proliferation of Karpas299 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 50 nM 200 nM 1,000 nM 5,000 nM 549148 99 106 106 89 609408 105 91 31 22 609416 104 119 72 59

TABLE 130 Dose-dependent reduction of proliferation of SupM2 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 16 nM 80 nM 400 nM 2,000 nM 549148 93 97 94 85 609408 93 96 55 3 609416 93 98 49 15

Example 27: Effect of Modified Oligonucleotides on Human IRF4 In Vitro, Multiple Doses

Modified oligonucleotides selected from the examples above were tested at various doses in mantle cell lymphoma (MCL) lines MAVER1, JVM2, Granta519, Mino, and Z138 for their effects on target knockdown and on cell line proliferation.

Target Knockdown

Cells were plated at a density of 10,000 cells per well and transfected by free uptake with 40 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. Control oligonucleotide 549148, a 3-10-3 cEt gapmer with the sequence GGCTACTACGCCGTCA (SEQ ID NO: 3385), was also included. After a treatment period of approximately 48 hours, RNA was isolated from the cells and IRF4 mRNA levels were measured by RT-qPCR. Human IRF4 primer probe set RTS4522 (described hereinabove in Example 11) was used to measure mRNA levels. IRF4 mRNA levels were adjusted according to total RNA content, as measured by RiboGreen. Results are presented as the percent level of IRF4 mRNA transcript, relative to untreated control (UTC) cells. As illustrated in the tables below, IRF4 mRNA levels were reduced in a dose-dependent manner in cells treated with modified oligonucleotides.

TABLE 131 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in MAVER1 cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 115 103 96 99 609408 121 102 80 35 609416 108 109 87 65

TABLE 132 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in JVM2 cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 108 114 108 100 609408 90 72 66 39 609416 101 82 74 93

TABLE 133 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in Granta519 cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 106 95 95 65 609408 176 165 134 122 609416 119 168 130 97 690890 161 160 143 102

TABLE 134 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in Mino cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 123 116 112 78 609408 124 109 67 31 609416 123 119 113 92 690890 122 116 103 42

TABLE 135 Dose-dependent reduction of IRF4 expression by modified oligonucleotides in Z138 cells Compound IRF4 expression (% UTC) Number 40 nM 200 nM 1,000 nM 5,000 nM 549148 68 72 78 19 609408 67 69 78 59 609416 76 69 89 72 690890 69 67 78 28 Proliferation Cells were plated at a density of 2,000 cells per well and transfected by free uptake with 50 nM, 200 nM, 1,000 nM, or 5,000 nM concentrations of modified oligonucleotide, as specified in the tables below. After seven days, CellTiterGlo-2.0 (Promega) was added and luminescence was measured on Glomax (Promega).

TABLE 136 Dose-dependent reduction of proliferation of MAVER1 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 80 nM 400 nM 2,000 nM 10,000 nM 549148 108 103 104 102 609408 104 102 83 40 609416 106 100 94 60

TABLE 137 Dose-dependent reduction of proliferation of JVM2 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 80 nM 400 nM 2,000 nM 10,000 nM 549148 108 114 108 100 609408 117 95 66 37 609416 114 101 97 86

TABLE 138 Dose-dependent reduction of proliferation of Granta519 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 80 nM 400 nM 2,000 nM 10,000 nM 549148 110 99 95 106 609408 34 10 4 2 609416 85 44 19 9 690890 58 24 12 7

TABLE 139 Dose-dependent reduction of proliferation of Mino cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 80 nM 400 nM 2,000 nM 10,000 nM 549148 90 103 106 101 609408 93 82 54 22 609416 94 84 68 56 690890 88 77 68 49

TABLE 140 Dose-dependent reduction of proliferation of Z138 cells by modified oligonucleotides Compound IRF4 proliferation (% UTC) Number 80 nM 400 nM 2,000 nM 10,000 nM 549148 91 88 89 82 609408 98 83 70 46 609416 91 82 74 56 690890 95 84 74 50

Example 28: In Vivo Activity in MM1.R Xenograft Model

A xenograft MM1.R model was used to evaluate activity of modified oligonucleotides targeted to human IRF4. Female NOD/SCID mice (JAX) at 4-6 weeks of age were given a subcuntaneous injection of 6 million MM1.R cells to form a xenograft tumor. Two weeks later, groups of 3 mice were administered 25, 50, or 100 mg/kg/dose modified oligonucleotide once a day for three days by subcutaneous injection. One group of mice received subcutaneous injections of PBS once a day for three days. The saline-injected group served as the control group to which oligonucleotide-treated groups were compared. Mice were sacrificed 48 hours after the final dose and tumors were collected for further analysis.

RNA Analysis

RNA was extracted from tumor tissue for RT-PCR analysis, which was performed as described above. Data were analyzed with primer probe set 34726, described above in Example 7 or primer probe set 35624 (forward sequence TCCCGTGTTGCTTCAAACT, designated herein as SEQ ID NO: 3395; reverse sequence TACCTGCTGGCAGTTCTTTC, designated herein as SEQ ID NO: 3396; probe sequence ACAGATGGGACTTAACAGGCAATGGG, designated herein as SEQ ID: 3397), which specifically detect human IRF4, as indicated in the tables below. Results are presented as percent change of mRNA, relative to PBS control, normalized with human B-Actin levels from the human tumor cells and mouse stromal cells using human specific primer probe set 5002 (forward sequence CGGACTATGACTTAGTTGCGTTAC; designated herein as SEQ ID NO: 3398; reverse sequence GCCATGCCAATCTCATCTTGT, designated herein as SEQ ID NO: 3399; probe sequence CCTTTCTTGACAAAACCTAACTTGCGCAGA, designated herein as SEQ ID NO: 3400).

TABLE 141 Activity of modified oligonucleotides in MM1.R xenograft model Compound IRF4 mRNA ID Dose PPset (% PBS) PBS n/a 34726 100 690890 25 34726 59 50 34726 53 100 34726 24 882800 50 34726 43 100 34726 18 935658 50 34726 52 100 34726 22 935918 50 34726 39 100 34726 18 935968 50 34726 52 100 34726 22 1012795 50 34726 45 100 34726 26 1014095 50 34726 71 100 34726 34 1014834 50 34726 56 100 34726 32 935762 50 34726 31 100 34726 27

TABLE 142 Activity of modified oligonucleotides in MM1.R xenograft model PBS n/a 34726 100 935696 50 34726 28 100 34726 13

TABLE 143 Activity of modified oligonucleotides in MM1.R xenograft model Compound IRF4 mRNA ID Dose Ppset (% PBS) PBS n/a 35624 100 690890 50 35624 55 100 35624 33 882800 50 35624 51 100 35624 31 935658 50 35624 56 100 35624 27 935918 50 35624 39 100 35624 9.9 935968 50 35624 26 100 35624 9.1 1012795 50 35624 18 100 35624 21 1014095 50 35624 28 100 35624 27 1014834 50 35624 33 100 35624 15 935762 50 35624 37 100 35624 22 935696 50 35624 19 100 35624 15 Protein Analysis Levels of hIRF4 protein were measured in the xenograft tumors by a human-specific IRF4 antibody (abcam EP5699) on the WES system (ProteinSimple). Levels of Igλ, a clinically-relevant biomarker for MM, were also measured on the WES system. Reductions of hIRF4 and Igλ were observed.

TABLE 144 Protein Levels in MM1.R Xenografts Compound Dose IRF4 Protein Igλ Protein ID (mg/kg/day) (% PBS) (% PBS) PBS n/a 100 100 690890 50 53 83 100 28 64 882800 50 67 88 100 27 68 935918 50 41 103 100 17 57 935968 50 36 91 100 15 52 1012795 50 32 105 100 14 60 1014095 50 35 78 100 15 56 1014834 50 57 83 100 31 69 935762 50 46 69 100 22 54 935696 50 26 40 100 15 37 935658 50 45 102 100 24 80

Example 29: Anti-Tumor Activity of Modified Oligonucleotides in a MM1.R Xenograft Model

A xenograft MM1.R model was used to evaluate activity of modified oligonucleotides targeted to human IRF4. Female NOD-SCID mice at 5-6 weeks of age were given a subcuntaneous injection of 3 million MM1.R cells to form a xenograft tumor. 23 days later, groups of 8 mice were administered 50 mg/kg/dose modified oligonucleotide five times a week by subcutaneous injection for 3.5 weeks. One group of mice received subcutaneous injections of PBS five times a week. The saline-injected group served as the control group to which oligonucleotide-treated groups were compared. Tumor volume was estimated by caliper measurement. Mice were sacrificed 24 hours after the last dose and tissue was collected for RNA and protein analysis.

Tumor Volume

TABLE 145 Tumor volume (mm³) Compound Days after MM1.R Cell Injection ID 23 27 30 34 37 41 44 PBS 215 374 590 936 1597  1877*  2530* 792169 228 400 615 1194  1451  1669*  2394* 1014834 225 319 451 606 931 1202  1733  1014095 222 291 504 554 886 795*  932* 1012795 221 355 459 472  610* n.d. n.d. 935968 210 313 463 587 771 447*  585* 935918 219 346 569 705 899 746* 1024* 935696 219 357 422 422 379 364  426 935658 269 340 557 619 856 983  1299  935762 220 290 397  488*  593* 770*  949* 882800 218 394 520 735 1075  907* 1279* 690890 216 307 419 498 643 830  1191  *Values represent the average of 3-7 mice Body Weight Body weights were measured throughout the study as a measure of tolerability.

TABLE 146 Body Weight (% of Day 23) Compound Days after MM1.R Cell Injection ID 23 27 30 34 37 41 44 PBS 100 101 101 106 104* 111* 115* 792169 100 106 106 116 121* 121* 126* 1014834 100 102 101 103 107  107  108  1014095 100 100 98 98  98*  98*  95* 1012795 100 101 97 82 n.d. n.d. n.d. 935968 100 102 101 103 102* 102* 101* 935918 100 101 101 102 100* 100*  98* 935762 100 102 99 103 104  104  100  935696 100 104 103 105 101  101  97 935658 100 100 100 100 98 98 95 882800 100 102 101 106 103* 103*  98* 690890 100 100 97 96 93 93 94 *Values represent the average of 3-7 animals Liver Function

To evaluate the effect of modified oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Plasma levels of ALT (alanine transaminase) were measured and the results are presented in the table below expressed in IU/L.

TABLE 147 Liver Transaminases Compound ID ALT (IU/L) PBS 24 792169 45 1014834 894 1014095 1601 1012795 1915 935968 289 935918 192 935762 1016 935696 1334 935658 91 882800 5219 690890 496 RNA and Protein Analysis

IRF4 mRNA in tumor samples was measured by RT-PCR using PPset RTS34726, described above. IRF4 protein in tumor samples was determined by western blot as described in Example 28 above.

TABLE 148 IRF4 Protein and mRNA Levels hIRF4 mRNA Level hIRF4 Protein Level PBS 98 100 792169 159 110 1014834 53 39 1014095 58 43 1012795 37 n.d. 935968 41 35 935918 28 31 935762 47 34 935696 39 27 935658 51 43 882800 57 32 690890 49 46

Example 30: Efficacy of Modified Oligonucleotides Targeted to hIRF4 in a Systemically Disseminated MM1.R Model with Bone Marrow Involvement

A systemically disseminated MM1.R model was used to evaluate activity of modified oligonucleotides targeted to human IRF4. Female nod-scid IL2Rγ^(null) mice at 4-6 weeks of age were first administered 50 mg/kg cyclophosphamide on day 0, and on day 1 were administrated 10 million MM1.R cells via an intravenous injection. On day 14, plasma human Igλ was tested by ELISA and mice were randomized to groups based on these results. Starting on day 21, groups of 4 mice were administered 50 mg/kg/day modified oligonucleotide once a day for three days, and sacrificed 48 hours after the last dose. Levels of hIRF4 mRNA were measured in bone marrow. The tumor burden was measured by measuring levels of hActin mRNA. Results are presented as percent change of mRNA, relative to PBS control treated mice.

TABLE 149 Bone marrow IRF4 mRNA and Tumor Burden. Compound IRF4 mRNA in bone marrow Tumor ID (% PBS) Burden PBS 100  4276 792169 70  830 882800 24  33 935918 36* 118 935696 38* 559 935968 34* 18 *Values represent the average of 1-3 mice, excluding mice with undetectable hIRF mRNA levels in bone marrow.

Example 31: Efficacy of Modified Oligonucleotides Targeted to hIRF4 in a Systemically Disseminated MM1.R Model with Bone Marrow Involvement

A systemically disseminated MM1.R model was used to evaluate activity of modified oligonucleotides targeted to human IRF4. Female NOD-SCID IL2Rγ^(null) mice at 4-6 weeks of age were first administered 50 mg/kg cyclophosphamide on day 0, and on day 1 were administrated 10 million MM1.R cells via an intravenous injection. On day 14, serum human Igλ was tested by ELISA and mice were randomized to groups based on these results. Starting on day 15, groups of ten mice were administered modified oligonucleotide with a loading dose of 50 mg/kg/day for 1 week, and then 3 doses a week at 50 mg/kg/day continuing until animal death, bodyweight drop of <20% or paralysis. One group of ten mice was administered PBS as a control, and another group was administered the control oligonucleotide 792169.

TABLE 150 Survival Percentage Treatment Compound ID Day PBS 792169 882800 935918 935696 935968 0 100 100 100 100 100 100 40 90 60 100 100 100 100 42 90 60 90 100 90 100 43 60 60 90 100 90 100 44 40 20 90 100 90 100 45 20 0 90 100 80 100 46 0 0 90 100 80 100 49 0 0 70 90 80 100 51 0 0 70 90 80 100

Example 32: Activity of Modified Oligonucleotides Targeting hIRF4 in a TMD8 Human ABC-DLBCL Tumor Model

A xenograft tumor model was used to evaluate activity of modified oligonucleotides targeted to human IRF4. 4 million ABC-DLBCL TMD8 cells were implanted into the flanks of 5 week old female NOD/SCID mice. When tumors reached an average volume of 100 mm³, approximately two weeks post-implantation, groups of eight mice were administered 50 mg/kg/day modified oligonucleotide for two weeks. Mice were sacrificed after the last dose and tumors were collected for mRNA analysis.

Tumor volume was estimated with caliper measurement. Levels of hIRF4 mRNA were measured in tumor tissue and normalized to control animals.

TABLE 151 Tumor volume (mm³) Days post-implantation Compound 15 19 22 25 27 29 32 ID Tumor Volume (mm³) PBS 112 276 602 872 1162 1365 2012 792169 112 232 488 797 1044 1359 1850 690890 114 206 392 481 589 675 777 882800 113 227 417 603 712 729 860 935696 113 225 448 570 676 761 729

TABLE 152 hIRF4 mRNA levels hIRF4 mRNA Level PBS 100 792169 127 690890 84 882800 72 935696 58

Example 33: Tolerability of Modified Oligonucleotides Targeting hIRF4 in Balb/c Mice

Balb/c mice are frequently utilized for safety and efficacy testing. The mice were treated with antisense oligonucleotides selected from studies described above and evaluated for changes in the levels of various plasma chemistry markers.

Treatment

Groups of 4-6 week old male Balb/c mice were injected subcutaneously twice a week for four weeks with 50 mg/kg of modified oligonucleotides (100 mg/kg/week dose). One group of male Balb/c mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis. Plasma Chemistry Markers To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of transaminases, bilirubin, and BUN were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). The results are presented in the table below. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.

TABLE 153 Plasma chemistry markers in Balb/c mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 37  84 22.9 0.25 609296 n.d. n.d. n.d. n.d. 609343 n.d. n.d. n.d. n.d. 609354 n.d. n.d. n.d. n.d. 609357 n.d. n.d. n.d. n.d. 609391 1799  1362  16.7 0.31 609408 859  507 18.4 0.28 609416 93 114 21.1 0.23 609296 n.d. n.d. n.d. n.d. 609547 982  802 27.3 0.34 609592 53 170 21.2 0.25

TABLE 154 Plasma chemistry markers in Balb/c mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 35.8 53 21.1 0.25 666431 85.5 120 21.1 0.29 666440 8173 3432 26.2 3.84 666449 150 124 24.9 0.18 666471 n.d. n.d. n.d. n.d. 666475 1989 1607 25.0 0.23 666496 2926 2423 22.4 0.30 666569 3427 3288 23.8 0.23 666575 525 385 24.5 0.21 666582 3543 2893 20.0 0.44 666584 43.3 57 25.4 0.21 666586 2970 2051 23.8 0.26 666587 1057 964 28.1 0.16 666649 4662 3970 23.8 10.99  666683 1363 1657 21.3 0.19

TABLE 155 Plasma chemistry markers in Balb/c mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS  32  89 24.1 0.29 609555 737 238 25.9 0.15 666772 n.d. n.d. n.d. n.d 666775 n.d. n.d. n.d. n.d 668818 n.d. n.d. n.d. n.d 668849 105 131 24.7 0.12 668850 547 394 27.8 0.11 668902  53 103 21.5 0.18 668934 n.d. n.d. n.d. n.d 668936 2260  3211  24.8 0.13 668937  39  89 23.2 0.14 668947 n.d. n.d. n.d. n.d 668998 102 101 21.7 0.14 668999 625 404 28.9 0.21 669018 198 157 28.1 0.17 669022  33  60 19.9 0.19 669040 176 170 23.2 0.11 669066 589 300 23.8 0.27 669067 n.d. n.d. n.d. n.d 669068  95 137 20.0 0.16 669075 647 400 19.4 0.15

TABLE 156 Plasma chemistry markers in Balb/c mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS  21  77 22.8 0.16 690508 n.d. n.d. n.d. n.d. 690509 n.d. n.d. n.d. n.d. 690510 n.d. n.d. n.d. n.d. 690511 n.d. n.d. n.d. n.d. 690512 n.d. n.d. n.d. n.d. 690514 595 440 19.3 0.15 690515 n.d. n.d. n.d. n.d. 690522 2440  1878  26.6 0.23 690527 n.d. n.d. n.d. n.d. 690861 933 752 12.2 0.19 690863 889 798 17.0 0.56 690865 n.d. n.d. n.d. n.d. 690873 1844  1160  16.6 0.28 690875 887 770 16.3 0.16 690877 n.d. n.d. n.d. n.d. 690879 887 652 22.2 0.21 690881 442 320 23.0 0.12 690883 1505  901 38.9 0.34 690890  76  64 25.6 0.14 690892  56 126 23.9 0.17 690898  47 120 27.4 0.11 691028 n.d. n.d. n.d. n.d. 691032 588 216 216   0.16 691033 709 451 31.4 0.13 Organ Weights Liver, kidney, and spleen weights were measured at the end of the study, and are presented as the percent change compared to PBS-treated animals in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.

TABLE 157 Organ Weights (g) Compound ID Liver Kidney Spleen PBS 1.25 0.39 0.11 609296 n.d. n.d. n.d. 609343 n.d. n.d. n.d. 609354 n.d. n.d. n.d. 609357 n.d. n.d. n.d. 609391 1.29 0.32 0.23 609408 1.48 0.37 0.14 609416 1.57 0.38 0.14 609296 n.d. n.d. n.d. 609547 2.56 0.35 0.55 609592 1.41 0.37 0.13

TABLE 158 Organ Weights (g) Compound ID Liver Kidney Spleen PBS 1.30 0.40 0.10 666431 1.21 0.38 0.15 666440 2.38 0.37 0.10 666449 1.39 0.40 0.11 666471 n.d. n.d. n.d. 666475 1.49 0.40 0.10 666496 1.66 0.44 0.12 666569 1.09 0.34 0.13 666575 1.33 0.37 0.10 666582 2.03 0.42 0.13 666584 1.34 0.40 0.11 666586 1.36 0.29 0.12 666587 1.77 0.38 0.11 666649 0.81 0.30 0.07 666683 2.05 0.38 0.12

TABLE 159 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 1.49 0.43 0.12 609555 1.50 0.36 0.17 666772 n.d. n.d. n.d. 666775 n.d. n.d. n.d. 668818 n.d. n.d. n.d. 668849 1.75 0.41 0.22 668850 1.96 0.41 0.16 668902 1.50 0.39 0.12 668934 n.d. n.d. n.d. 668936 1.97 0.33 0.13 668937 1.58 0.41 0.14 668947 n.d. n.d. n.d. 668998 1.53 0.38 0.16 668999 1.64 0.38 0.13 669018 1.92 0.41 0.14 669022 1.47 0.41 0.12 669040 1.66 0.44 0.11 669066 1.64 0.39 0.15 669067 1.52 0.38 0.13 669068 n.d. n.d. n.d. 669075 1.43 0.36 0.17

TABLE 160 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 0.86 0.29 0.10 690508 n.d. n.d. n.d. 690509 n.d. n.d. n.d. 690510 n.d. n.d. n.d. 690511 n.d. n.d. n.d. 690512 n.d. n.d. n.d. 690514 1.68 0.28 0.19 690515 n.d. n.d. n.d. 690522 1.40 0.26 0.11 690527 n.d. n.d. n.d. 690861 1.29 0.27 0.12 690863 1.27 0.27 0.15 690865 n.d. n.d. n.d. 690873 1.17 0.28 0.14 690875 1.36 0.28 0.17 690877 n.d. n.d. n.d. 690879 1.04 0.26 0.14 690881 1.19 0.28 0.13 690883 1.59 0.30 0.10 690890 0.97 0.27 0.13 690892 1.08 0.30 0.17 690898 1.08 0.30 0.15 691028 n.d. n.d. n.d. 691032 1.33 0.29  0.1525 691033  1.625  0.3175  0.125

Example 34: Tolerability of Modified Oligonucleotides Targeting hIRF4 in CD1 Mice

CD1® mice (Charles River, Mass.) are frequently utilized for safety and efficacy testing. The mice were treated with antisense oligonucleotides selected from studies described above and evaluated for changes in the levels of various plasma chemistry markers.

Treatment

Groups of 4-6 week old male CD1 mice were injected subcutaneously twice a week for four weeks with 50 mg/kg of modified oligonucleotides (100 mg/kg/week dose). One group of male CD1 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis. Plasma Chemistry Markers To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of transaminases, bilirubin, and BUN were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). The results are presented in the table below. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.

TABLE 161 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 27 65 19.7 0.19 881413 43 78 19.3 0.18 881442 594 484 23.1 0.24 881449 36 52 19.9 0.24 881450 100 146 20.8 0.18 881506 683 417  8.7 0.18 881517 121 103 22.1 0.16 881581 501 413 23.7 0.27 881610 44 79 18.5 0.19 881658 39 66 16.0 0.17 881659 4924 3485 15.3 5.48 881660 267 176 19.7 0.16 881728 644 389 21.1 0.23 881742 780 443 17.6 0.23 882099 3607 1971 23.1 0.28 882282 n.d. n.d. n.d. n.d. 882305 2688 1379 24.9 0.31 882433 2683 1944 19.5 0.42

TABLE 162 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 23 69 17.0 0.19 881449 34 65 19.2 0.23 881582 206 211 18.6 0.15 881588 902 829 15.2 0.32 881658 334 226 14.0 0.18 881727 1120 748 16.8 0.40 792169 23 43 18.0 0.21

TABLE 163 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 25  41 22.2 0.17 935765 n.d. n.d. n.d. n.d. 935580 n.d. n.d. n.d. n.d. 935805 n.d. n.d. n.d. n.d. 935853 n.d. n.d. n.d. n.d. 935824 n.d. n.d. n.d. n.d. 935581 n.d. n.d. n.d. n.d. 935689 n.d. n.d. n.d. n.d. 935698 1462 1523  17.5 0.19 935699 n.d. n.d. n.d. n.d. 935707 1334 703 17.3 0.21 935679 n.d. n.d. n.d. n.d. 935727 n.d. n.d. n.d. n.d. 935795 n.d. n.d. n.d. n.d. 935898 101 129 20.3 0.18 935782 680 278 19.0 0.20 935878 1805 1128  19.1 0.21 935850 661 492 21.2 0.08 935724 1630 1341  19.9 0.25 935658 53  65 19.5 0.24 935762 106 132 18.0 0.18 935851 513 300 18.0 0.13

TABLE 164 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 19 34 18.6 0.17 935921 708 372 15.5 0.19 935961 1068 754 14.8 0.28 936018 609 358 17.9 0.14 936039 n.d. n.d. n.d. n.d. 935708 n.d. n.d. n.d. n.d. 935958 97 117 17.5 0.10 935854 304 205 18.6 0.14 935968 53 70 18.9 0.12 935620 1883 2950 17.2 1.17 935697 1504 763 16.9 0.22 935700 722 480 18.8 0.18 935734 1253 500 17.9 0.17 935857 1270 1233 17.7 0.29 936016 731 571 22.7 0.15 936006 38 47 16.9 0.13 936007 131 152 18.1 0.10 935948 192 183 19.5 0.15 935603 193 260 17.0 0.19 935701 426 277 18.0 0.06 935928 69 84 17.4 0.10

TABLE 165 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 23 44 22.5 0.20 969991 n.d. n.d. n.d. n.d. 970013 n.d. n.d. n.d. n.d. 970043 636 610 19.6 0.33 970103 786 512 20.1 0.17 970117 1786 877 22.5 0.29 970139 1011 1244 21.0 1.69 970159 1772 3821 15.7 2.68 970162 470 422 18.0 0.15 970189 353 377 21.0 0.19 970212 1714 1062 22.7 1.00 970524 n.d. n.d. n.d. n.d. 970527 182 181 18.1 0.12

TABLE 166 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 24 99 20.1 0.21 881955 4127 3743 45.7 2.61 882085 53 73 22.3 0.18 882086 2522 2002 25.5 0.43 882087 377 276 23.1 0.15 882175 575 554  9.4 0.68 882177 2666 3030 26.6 4.87 882228 893 1659 19.3 0.22 882398 980 1070 22.1 7.91 882408 46 69 17.9 0.17 882800 70 84 23.4 0.20 969933 383 272 21.7 0.15 969936 n.d. n.d. n.d. n.d. 969938 1142 1009 18.1 0.76 970211 301 224 20.1 0.15 970249 1724 2104 20.0 6.21 970545 836 1049 24.8 0.34 970546 1248 1334 23.8 0.31 970547 878 665 18.4 0.53 970548 1607 3352 17.7 4.86 970552 n.d. n.d. n.d. n.d.

TABLE 167 Plasma chemistry markers in CD1 mouse plasma at week 4 Compound ALT AST BUN T. Bil ID (U/L) (U/L) (mg/dL) (mg/dL) PBS 23 44 19.7 0.22 1012795 143 235 20.9 0.16 1012821 1189 1434 24.6 0.22 1012884 1523 1939 27.0 0.69 1014095 63 71 19.4 0.18 1014393 666 627 19.3 0.25 1014394 117 129 17.8 0.16 1014834 301 190 19.7 0.21 1015716 230 205 18.4 0.17

TABLE 168 Plasma chemistry markers in CD1 mice plasma at week 4 Compound ALT AST T. Bil BUN ID (U/L) (U/L) (mg/dL) (mg/dL) Albumin PBS 21 45 0.190 21.8 2.54 935918 87 91 0.118 18.8 2.07 935696 71 108 0.143 21.7 2.22 Organ Weights Liver, kidney, and spleen weights were measured at the end of the study, and are presented as the percent change compared to PBS-treated animals in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.

TABLE 169 Organ Weights (g) Compound ID Liver Kidney Spleen PBS 2.1 0.56 0.141 881413 2.2 0.57 0.217 881442 2.7 0.52 0.186 881449 2.1 0.47 0.169 881450 2.2 0.50 0.151 881506 2.9 0.62 0.191 881517 2.4 0.48 0.180 881581 2.2 0.51 0.192 881610 1.9 0.53 0.165 881658 2.0 0.54 0.158 881659 1.7 0.43 0.153 881660 1.9 0.52 0.175 881728 2.3 0.48 0.161 881742 1.9 0.48 0.127 882099 2.2 0.47 0.193 882282 n.d. n.d. n.d. 882305 2.1 0.52 0.181 882433 2.0 0.54 0.155

TABLE 170 Organ weights (g) Compound ID Liver Kidney Spleen PBS 1.8 0.54 0.111 881449 2.1 0.54 0.146 881582 2.0 0.48 0.177 881588 1.9 0.39 0.171 881658 2.2 0.58 0.188 881727 2.1 0.51 0.201 792169 1.9 0.55 0.128 UTC 2.0 0.69 0.144

TABLE 171 Organ weights (g) Compound ID Liver Kidney Spleen PBS 2.0 0.50 0.109 935765 n.d. n.d. n.d. 935580 n.d. n.d. n.d. 935805 n.d. n.d. n.d. 935853 n.d. n.d. n.d. 935824 n.d. n.d. n.d. 935581 n.d. n.d. n.d. 935689 n.d. n.d. n.d. 935698 2.4 0.55 0.213 935699 n.d. n.d. n.d. 935707 2.3 0.54 0.199 935679 n.d. n.d. n.d. 935727 n.d. n.d. n.d. 935795 n.d. n.d. n.d. 935898 2.2 0.58 0.245 935782 2.9 0.56 0.279 935878 2.7 0.56 0.192 935850 2.8 0.70 0.203 935724 2.6 0.45 0.110 935658 2.0 0.71 0.205 935762 2.3 0.57 0.197 935851 2.4 0.60 0.176

TABLE 172 Organ weights (g) Compound ID Liver Kidney Spleen PBS 1.9 0.52 0.117 935921 2.3 0.51 0.219 935961 2.1 0.53 0.190 936018 2.2 0.41 0.200 936039 n.d. n.d. n.d. 935708 n.d. n.d. n.d. 935958 2.2 0.47 0.219 935854 2.0 0.44 0.102 935968 2.4 0.58 0.157 935620 3.2 0.57 0.243 935697 2.9 0.56 0.208 935700 2.0 0.49 0.121 935734 2.7 0.55 0.162 935857 1.5 0.34 0.110 936016 2.8 0.57 0.416 936006 2.3 0.48 0.136 936007 2.3 0.55 0.149 935948 1.9 0.54 0.166 935603 1.6 0.49 0.125 935701 1.9 0.43 0.144 935928 2.3 0.53 0.155

TABLE 173 Organ weights (g) Compound ID Liver Kidney Spleen PBS 1.8 0.51 0.133 969991 n.d. n.d. n.d. 970013 n.d. n.d. n.d. 970043 2.3 0.48 0.238 970103 2.2 0.54 0.175 970117 3.2 0.48 0.255 970139 2.7 0.56 0.210 970159 2.7 0.62 0.183 970162 2.2 0.54 0.215 970189 3.1 0.45 0.135 970212 2.3 0.45 0.218 970524 n.d. n.d. n.d. 970527 2.4 0.60 0.178

TABLE 174 Organ weights (g) Compound ID Liver Kidney Spleen PBS 1.7 0.42 0.095 881955 1.8 0.32 0.110 882085 2.0 0.45 0.110 882086 4.9 0.41 0.258 882087 2.4 0.47 0.130 882175 1.4 0.46 0.098 882177 3.6 0.43 0.185 882228 2.8 0.49 0.193 882398 1.7 0.44 0.113 882408 2.0 0.51 0.135 882800 1.9 0.41 0.128 969933 2.2 0.50 0.145 969936 n.d. n.d. n.d. 969938 1.8 0.45 0.150 970211 2.1 0.40 0.143 970249 3.3 0.65 0.190 970545 2.5 0.44 0.133 970546 0.9 0.28 0.060 970547 2.1 0.50 0.243 970548 2.8 0.52 0.260 970552 n.d. n.d. n.d.

TABLE 175 Organ weights (g) Compound Liver Kidney Spleen PBS 1.78 0.56 0.115 935918 2.44 0.62 0.167 935696 2.20 0.61 0.352

Example 35: Tolerability of Modified Oligonucleotides Targeting hIRF4 in Sprague-Dawley Rats

Sprague-Dawley rats are a multipurpose model used for safety and efficacy evaluations. The rats were treated with modified antisense oligonucleotides from the studies described in the Examples above and evaluated for changes in the levels of various plasma chemistry markers.

Treatment

Male Sprague-Dawley rats were maintained on a 12-hour light/dark cycle and fed ad libitum with Purina normal rat chow, diet 5001. Groups of 4Sprague-Dawley rats each were injected subcutaneously once a week for 6 weeks with 50 mg/kg of ISIS oligonucleotide (50 mg/kg weekly dose). Forty eight hours after the last dose, rats were euthanized and organs and plasma were harvested for further analysis.

Liver and Kidney Function

To evaluate the effect of modified oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Plasma levels of ALT (alanine transaminase), AST (aspartate transaminase), blood urea nitrogen (BUN), and T. bilirubin were measured and the results are presented in the table below. Plasma levels of bilirubin were also measured using the same clinical chemistry analyzer and the results are also presented in the table below. Values represent the % change normalized to PBS-treated animals. Modified oligonucleotides that caused changes in the levels of any markers of liver function outside the expected range for antisense oligonucleotides were excluded in further studies.

TABLE 176 Liver function markers in Sprague-Dawley rats Compound ALT AST T. Bil BUN Albumin ID (IU/L) (IU/L) (mg/dL) (mg/dL) (g/dL) PBS 47 67 0.14 16.4 3.92 690898 41 73 0.17 20.7 3.19 881413 42 61 0.18 18.1 3.66 881449 47 67 0.26 19.7 3.77 935658 47 55 0.16 17.4 3.80 935696 41 70 0.18 16.9 3.55 935898 72 81 0.19 18.0 4.23 935928 189 188 0.19 19.7 3.59 935968 39 85 0.21 17.2 3.61 936006 40 55 0.14 14.3 3.97

TABLE 177 Liver function markers in Sprague-Dawley rats Compound ALT AST T. Bil BUN Albumin ID (IU/L) (IU/L) (mg/dL) (mg/dL) (g/dL) PBS 34 58 0.09 15.7 2.20 690890 32 70 0.16 20.1 2.39 690892 42 77 0.15 18.5 1.82 882085 68 73 0.09 47.1 1.36 882800 40 100 0.16 20.2 2.30 970527 45 80 0.12 19.8 2.09

TABLE 178 Liver function markers in Sprague-Dawley rats Compound ALT AST T. Bil BUN Albumin ID (IU/L) (IU/L) (mg/dL) (mg/dL) (g/dL) PBS 41 70 0.14 14.4 3.60 882408 38 68 0.15 17.0 2.80 1012795 25 70 0.09 22.3 2.47 1014095 37 101 0.12 20.3 2.41 1014393 41 70 0.09 20.3 2.28 1014394 32 74 0.08 21.1 2.53 1014834 34 68 0.10 14.2 3.04

TABLE 179 Liver function markers in Sprague-Dawley rats Compound ALT AST T. Bil BUN Albumin ID (IU/L) (IU/L) (mg/dL) (mg/dL) (g/dL) PBS 55 51 0.15 17.1 3.04 935762 35 53 0.14 18.7 2.63 935918 31 55 0.15 19.7 2.60 Hematology Assays

Blood obtained from all rat groups were sent to Antech Diagnostics for hematocrit (HCT) measurements and analysis, as well as measurements of the various blood cells, such as WBC, RBC, and total hemoglobin content. The results are presented in the table below. Modified oligonucleotides that caused changes in the levels of any of the hematology markers outside the expected range for antisense oligonucleotides were excluded in further studies.

TABLE 180 Hematology markers in Sprague-Dawley rats Compound WBC RBC HGB HCT LYM MON EOS BAS PLT Retic ID (K/μL) (M/μL) (g/dL) (%) (/μL) (/μL) (/μL) (/μL) (K/μL) (K/μL) PBS 10.38 8.15 14.80 43.9 8711 357 68.3 12.3 868 252 690898 17.83 6.46 12.27 37.1 15745 1109 75.0 145.0 214 201 881413 7.08 6.71 12.55 36.6 6342 286 44.3 15.3 523 152 881449 6.68 7.53 13.60 39.8 5414 396 153.0 27.5 583 133 935658 9.73 7.97 14.53 42.4 8250 777 19.7 70.0 685 138 935696 7.80 7.33 13.10 38.6 6354 645 48.8 23.0 480 97 935898 7.75 8.64 15.20 43.3 6248 529 29.3 26.8 712 114 935928 13.15 7.36 13.23 40.1 7231 1177 8.5 90.3 476 160 935968 6.15 6.17 11.28 33.7 5292 436 8.0 68.8 462 178 936006 7.28 8.07 14.45 42.7 5875 729 20.3 57.3 621 220

TABLE 181 Hematology markers in Sprague-Dawley rats Compound WBC RBC HGB HCT LYM MON EOS BAS PLT Retic ID (K/μL) (M/μL) (g/dL) (%) (/μL) (/μL) (/μL) (/μL) (K/μL) (K/μL) PBS 4.35 5.18 9.68 29.4 3546 116 102.8 10.5 292 136 690890 4.75 4.84 8.98 27.4 3844 325 25.3 55.0 190 113 690892 4.70 5.46 10.65 31.4 3917 276 42.3 34.5 184 120 882085 7.18 5.18 9.43 29.2 5159 758 73.0 24.5 352 122 882800 5.10 5.08 9.13 27.9 4190 479 46.0 40.3 165 157 970527 9.60 6.10 10.88 32.4 7990 846 28.3 50.8 249 123

TABLE 182 Hematology markers in Sprague-Dawley rats Compound WBC RBC HGB HCT LYM MON EOS BAS PLT Retic ID (K/μL) (M/μL) (g/dL) (%) (/μL) (/μL) (/μL) (/μL) (K/μL) (K/μL) PBS 9.35 7.81 14.28 46.6 7820 330 208.3 11.3 344 214 882408 13.25 9.46 16.73 52.0 8387 1020 35.3 29.3 407 163 1012795 15.20 4.86 10.03 33.6 11767 1996 28.5 12.8 149 304 1014095 18.55 6.93 12.35 41.4 11958 1500 95.0 22.3 727 281 1014393 21.90 7.25 13.08 41.8 17709 2038 50.5 156.3 491 123 1014394 23.55 7.21 13.58 43.3 19656 2121 31.8 67.5 270 207 1014834 14.43 7.97 14.65 45.9 12543 1009 42.3 180.0 395 182

TABLE 183 Hematology markers in Sprague-Dawley rats Compound WBC RBC HGB HCT LYM MON EOS BAS PLT Retic ID (K/μL) (M/μL) (g/dL) (%) (/μL) (/μL) (/μL) (/μL) (K/μL) (K/μL) PBS 11.13 9.45 16.70 52.8 9156 686 103 8.0 82 238 935762 14.53 7.72 13.43 42.4 13586 510 0 29.3 92 232 935918 9.25 7.08 12.50 40.5 8509 456 51 5.0 92 286 Organ Weights

Liver, heart, spleen and kidney weights were measured at the end of the study, and are presented in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for antisense oligonucleotides were excluded from further studies.

TABLE 184 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 15.8 2.91 0.90 690898 16.9 3.18 3.65 881413 15.0 2.83 1.32 881449 15.4 3.07 1.19 935658 13.8 2.40 1.33 935696 12.6 2.60 1.22 935898 12.0 2.44 1.07 935928 16.8 3.23 1.94 935968 14.9 2.85 1.56 936006 14.2 2.75 1.59

TABLE 185 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 15.2 3.38 0.72 690890 15.8 3.17 1.62 690892 15.2 3.27 1.72 882085 12.5 3.78 1.41 882800 17.0 3.46 3.24 970527 14.0 3.72 1.94

TABLE 186 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 18.6 3.70 0.92 882408 15.1 3.60 1.51 1012795 18.9 3.70 4.06 1014095 18.2 3.63 3.05 1014393 15.9 3.98 2.30 1014394 18.3 4.41 2.79 1014834 14.4 3.35 5.01

TABLE 187 Organ weights (g) Compound ID Liver (g) Kidney (g) Spleen (g) PBS 14.6 3.20 0.88 935762 16.1 3.43 1.73 935918 18.7 3.89 2.01

Example 36: Tolerability of Modified Oligonucleotides in Non-Human Primates

Modified oligonucleotides described above were further evaluated for potency in non-human primates.

Treatment

Male cynomolgus monkeys were divided into groups of 4 non-human primates (NHP) each. Groups received a dose of 40 mg/kg of modified oligonucleotide by subcutaneous injection on day 1, 3, 5, and 7, and then once/week for six weeks. One group of NHP received doses of PBS. The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared. After six weeks, NHP were sacrificed and tissues were collected for analysis.

Tolerability

To evaluate the effect of these antisense oligonucleotides on liver and kidney function, samples of blood, plasma, serum and urine were collected from all study groups on day 44. The blood samples were collected via femoral venipuncture, 48 hrs post-dosing. The monkeys were fasted overnight prior to blood collection. Approximately 1.5 mL of blood was collected from each animal into tubes without anticoagulant for serum separation. Levels of the various markers were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Total urine protein and urine creatinine levels were measured, and the ratio of total urine protein to creatinine (P/C Ratio) was determined.

To evaluate the effect of the antisense oligonucleotides on hepatic function, plasma concentrations of transaminases (ALT, AST), Albumin (Alb) and total bilirubin (“T. Bil”) were measured. To evaluate the effect of the antisense oligonucleotides on kidney function, plasma concentrations of blood urea nitrogen (BUN) and creatinine (Cre) were measured. Urine levels of albumin (Alb), creatinine (Cre) and total urine protein (Micro Total Protein (MTP)) were measured, and the ratio of total urine protein to creatinine (P/C ratio) was determined.

To evaluate any inflammatory effect of the ISIS oligonucleotides in cynomolgus monkeys, C-reactive protein (CRP), which is synthesized in the liver and which serves as a marker of inflammation, was measured on day 44. For this, blood samples were taken from fasted monkeys, the tubes were kept at room temperature for a minimum of 90 min., and centrifuged at 3,000 rpm for 10 min at room temperature to obtain serum. The results are presented in the Tables below and indicate that most of the antisense oligonucleotides targeting human IRF4 were well tolerated in cynomolgus monkeys. ION 935918 and 935968 were well tolerated.

TABLE 188 Serum and urine clinical chemistry Serum (day 44) Urine (day 42) ISIS C3 ALT AST Alb BUN CRP Cre T. bil Alb Cre P/C No. mg/dL U/L U/L g/dL mg/dL mg/L mg/dL mg/dL g/dL mg/dL ratio PBS 120.73 48.00 69.10 4.35 25.60 1.66 0.858 0.29 0.35 55.0 0.00 690890 71.20 50.18 75.20 3.66 26.10 9.52 0.988 0.23 0.53 56.2 0.04 935658 95.90 42.33 81.85 3.99 21.70 4.53 0.808 0.29 0.68 39.0 0.12 935696 88.58 48.32 62.78 4.03 26.06 6.55 0.780 0.26 0.18 36.6 0.04 935762 100.10 47.58 52.63 3.91 24.98 6.88 0.76 0.25 0.03 37.9 0.01 935918 94.45 49.45 79.20 4.05 29.85 4.65 0.90 0.24 0.12 60.2 0.01 935968 108.83 54.45 83.33 4.10 28.68 6.65 0.813 0.23 0.10 38.8 0.07 882800 100.83 46.55 61.80 4.00 24.10 2.53 0.818 0.22 0.36 34.9 0.14 1012795 82.05 37.08 51.40 3.05 27.45 21.28 0.763 0.20 0.50 55.0 0.10 1014095 85.83 38.15 46.85 2.81 15.10 11.16 0.593 0.15 0.03 43.9 0.07 1014834 89.13 54.73 75.28 3.88 23.65 7.03 0.978 0.25 0.14 75.9 0.05

TABLE 189 Body Weight Compound ID Body Weight (g) day 1 Body weight (g) day 42 PBS 2521 2594 690890 2508 2514 935658 2499 2557 935696 2412 2511 935762 2473 2653 935918 2483 2657 935968 2605 2798 882800 2577 2676 1012795 2623 2577 1014095 2567 2719 1014834 2556 2685 RNA Analysis

RNA was extracted from various tissues for real-time PCR analysis of mRNA expression of IRF4 as in previous examples. Results are presented as percent change of mRNA, relative to PBS control, normalized with NHP Cyclophylin A. As shown in the table below, treatment with modified oligonucleotides resulted in reduction of IRF4 mRNA in comparison to the PBS control with some of the treatment groups.

TABLE 190 % Inhibition of cynomolgus IRF4 Compound ID Bone marrow PBMC Spleen PBS 100 100 100 690890* 98 119 205 935658* 174 170 200 935696* 71 80 125 935762* 112 90 129 935918* 98 41 192 935968* 129 49 131  882800*** 107 75 133 1012795*  97 95 151  1014095*** 80 80 188 1014834*  139 180 192 *Compounds have one mismatch to cynomolgus monkey IRF4; ***compounds have three mismatches to cynomolgus monkey IRF4.

Example 37: Viscosity

Viscosity of modified oligonucleotide solutions was measured. The viscosity of 935918 is compatible with weekly subcutaneous injection, and the viscosities of both 935918 and 935968 are compatible with IV dosing.

TABLE 191 Viscosity Compound ID Dose (mg/mL) by weight Viscocity (cP) 690890 300 16.14 935658 300 48.79 935696 300 120.2 935762 300 11.39 935918 100 2.12 935968 300 53.76 882800 100 3.4 

What is claimed:
 1. A modified oligonucleotide according to the following chemical structure:


2. A composition comprising the modified oligonucleotide of claim 1 and a pharmaceutically acceptable carrier.
 3. The composition of claim 2, wherein the pharmaceutically acceptable carrier is water.
 4. The composition of claim 2, wherein the pharmaceutically acceptable carrier is phosphate buffered saline (PBS).
 5. A method of treating cancer in an individual comprising administering to the individual the compound of claim
 1. 6. The method of claim 5, wherein the cancer is a blood cancer, myeloma, multiple myeloma, B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia.
 7. The method of claim 6, wherein the cancer is multiple myeloma.
 8. The method of claim 5, wherein administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 9. The method of claim 6, wherein administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 10. The method of claim 7, wherein administering the compound inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 11. A method of treating cancer in an individual comprising administering to the individual the composition of claim
 2. 12. The method of claim 11, wherein the cancer is a blood cancer, myeloma, multiple myeloma, B cell malignancy, lymphoma, B cell lymphoma, T cell lymphoma, or leukemia.
 13. The method of claim 12, wherein the cancer is multiple myeloma.
 14. The method of claim 11, wherein administering the composition inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 15. The method of claim 12, wherein administering the composition inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 16. The method of claim 13, wherein administering the composition inhibits or reduces cancer cell proliferation, tumor growth, or metastasis.
 17. The method of claim 5, wherein the compound is administered parenterally.
 18. The method of claim 11, wherein the composition is administered parenterally.
 19. A method of inhibiting expression of IRF4 in a cancer cell comprising contacting the cancer cell with the compound of claim
 1. 20. The method of claim 19, wherein the cancer cell is a bone marrow cell, lymphoid cell, or lymph node cell.
 21. A method of inhibiting expression of IRF4 in a cancer cell comprising contacting the cancer cell with the composition of claim
 2. 22. The method of claim 21, wherein the cancer cell is a bone marrow cell, lymphoid cell, or lymph node cell. 